SEARCH · PubMed Health

Results for “Atomic force microscopy (AFM)”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

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

At least 271 records · Page 15Linked to original sources

PubMed1999

AFM study of citric acid-ferric chloride etching characteristics of dentin.

PURPOSE: To determine the etching characteristics of 10% citric acid etchant with 3% ferric chloride (10-3) compared with related etchants without ferric chloride (10-0), or one of similar pH (36% citric acid). MATERIALS AND METHODS: Atomic force microscopy (AFM) was used to measure the surface recession and morphology of dentin. AFM images were taken of the same samples at seven steps of an etching and drying procedure using 10-3 (pH = 1.09), 10-0 (pH = 1.87) or 36% (pH = 1.08) citric acid solutions: at baseline, after etching 15 s, after 5-s air-drying, after rewetting, after 24-hr desiccation, after 30 s rewetting and after 24 hrs rewetting in filtered and purified water. RESULTS: 10-3 etched surfaces receded more than either 10-0 or 36% etched surfaces, and underwent greater collapse on 5-s air-drying. On rewetting all surfaces re-expanded to near their as-etched level. Following prolonged desiccation (24 hrs) all samples collapsed to the level seen after brief air-drying. 30-s rewetting did not restore the surface, but re-expansion of the collapsed collagen matrix was seen at 24 hrs.

Acid Etching, Dental↗

PubMed2004

Comparison of antibody--antigen interactions on collagen measured by conventional immunological techniques and atomic force microscopy.

We have developed a means of using atomic force microscopy (AFM) to repeatedly localize a small area of interest (4 x 4 microm(2)) within a 0.5-cm(2) area on a heterogeneous sample, to obtain and localize high-resolution images and force measurements on nonideal samples (i.e., samples that better reflect actual biological systems, not prepared on atomically flat surfaces). We demonstrate the repeated localization and measurement of unbinding forces associated with antibody--antigen (ab--ag) interactions, by applying AFM in air and in liquid to visualize and measure polyclonal ab--ag interactions, using chicken collagen as a model system. We demonstrate that molecular interactions, in the form of ab--ag complexes, can be visualized by AFM when secondary antibodies are conjugated to 20-nm colloidal gold particles. We then compare those results with established immunological techniques, to demonstrate broader application of AFM technology to other systems. Data from AFM studies are compared with results obtained using immunological methods traditionally employed to investigate ab--ag interactions, including enzyme-linked immunosorbent assay, immunoblotting, and in situ immunofluorescence. Finally, using functionalized AFM tips with a flexible tether [poly(ethylene glycol) 800] to which a derivatized antibody was attached, we analyzed force curve data to measure the unbinding force of collagen antibody from its antigen, obtaining a value of approximately 90 +/- 40 pN with a MatLab code written to automate the analyses of force curves obtained in force--volume mode. The methodology we developed for embedded collagen sections can be readily applied to the investigation of other receptor--ligand interactions.

Air↗

PubMed2005

Frequency modulation atomic force microscopy reveals individual intermediates associated with each unfolded I27 titin domain.

In this study, we apply a dynamic atomic force microscopy (AFM) technique, frequency modulation (FM) detection, to the mechanical unfolding of single titin I27 domains and make comparisons with measurements made using the AFM contact or static mode method. Static mode measurements revealed the well-known force transition occurring at 100-120 pN in the first unfolding peak, which was less clear, or more often absent, in the subsequent unfolding peaks. In contrast, some FM-AFM curves clearly resolved a force transition associated with each of the unfolding peaks irrespective of the number of observed unfolded domains. As expected for FM-AFM, the frequency shift response of the main unfolding peaks and their intermediates could only be detected when the oscillation amplitudes used were smaller than the interaction lengths being measured. It was also shown that the forces measured for the dynamical interaction of the FM-AFM technique were significantly lower than those measured using the static mode. This study highlights the potential for using dynamic AFM for investigating biological interactions, including protein unfolding and the detection of novel unfolding intermediates.

Aluminum Silicates↗

PubMedDate not supplied

The interaction of DNA with bacteriophage phi 29 connector: a study by AFM and TEM.

The connector of bacteriophage phi 29 is involved in DNA packaging during viral morphogenesis and we have studied its in vitro binding to DNA using either linear or circular DNA. The protein-DNA complexes have been analyzed by transmission electron microscopy (TEM) and by atomic force microscopy (AFM) of samples directly deposited on mica. TEM showed the presence of a specific binding due to the interaction of the protein with the free ends of the DNA. The study of these samples by AFM showed two major types of morphologies: The interaction of the connector with circular DNA revealed that the strands of DNA that enter and exit the protein complex form an angle with a mean value of 132 degrees. Nevertheless, when the connector was incubated with linear DNA (and later circularized), there was an additional bend angle of about 168 degrees. Further morphological analysis of the latter samples by AFM revealed a structure of the protein-DNA complex consistent with the DNA traversing the connector, probably through the inner channel. On the other hand, images from the samples obtained by incubation of the connector with circular DNa were consistent with an interaction of the DNA with the outer side of the connector.

Bacillus Phages↗

PubMedDate not supplied

Role of ionic strength on the relationship of biopolymer conformation, DLVO contributions, and steric interactions to bioadhesion of Pseudomonas putida KT2442.

Biopolymers produced extracellularly by Pseudomonas putida KT2442 were examined via atomic force microscopy (AFM) and single molecule force spectroscopy. Surface biopolymers were probed in solutions with added salt concentrations ranging from that of pure water to 1 M KCl. By studying the physicochemical properties of the polymers over this range of salt concentrations, we observed a transition in the steric and electrostatic properties and in the conformation of the biopolymers that were each directly related to bioadhesion. In low salt solutions, the electrophoretic mobility of the bacterium was negative, and large theoretical energy barriers to adhesion were predicted from soft-particle DLVO theory calculations. The brush layer in low salt solution was extended due to electrostatic repulsion, and therefore, steric repulsion was also high (polymers extended 440 nm from surface in pure water). The extended polymer brush layer was "soft", characterized by the slope of the compliance region of the AFM approach curves (-0.014 nN/nm). These properties resulted in low adhesion between biopolymers and the silicon nitride AFM tip. As the salt concentration increased to > or =0.01 M, a transition was observed toward a more rigid and compressed polymer brush layer, and the adhesion forces increased. In 1 M KCl, the polymer brush extended 120 nm from the surface and the rigidity of the outer cell surface was greater (slope of the compliance region = -0.114 nN/nm). A compressed and more rigid polymer layer, as well as a less negative electrophoretic mobility for the bacterium, resulted in higher adhesion forces between the biopolymers and the AFM tip. Scaling theories for polyelectrolyte brushes were also used to explain the behavior of the biopolymer brush layer as a function of salt concentration.

Bacterial Adhesion↗

PubMed1994

Longitudinal patterns similar to G-banding in untreated human chromosomes: evidence from atomic force microscopy.

The structure of human metaphase chromosomes, fixed according to standard procedures for optical microscopy but not treated for banding, was examined by atomic force microscopy (AFM). The images show that chromosomes display a banding pattern very similar to G-banding, detected by the AFM as a variation in the thickness of chromatin. This similarity allows the identification of individual chromosomes.

Chromatin↗

PubMed1999

Charting the surfaces of the purple membrane.

The preponderance of structural data of the purple membrane from X-ray diffraction (XRD), electron crystallography (EC), and atomic force microscopy (AFM) allows us to ask questions about the structure of bacteriorhodopsin itself, as well as about the information derived from the different techniques. The transmembrane helices of bacteriorhodopsin are quite similar in both EC and XRD models. In contrast, the loops at the surfaces of the purple membrane show the highest variability between the atomic models, comparable to the height variance measured by AFM. The excellent agreement of the AFM topographs with the atomic models from XRD builds confidence in the results. Small technical difficulties in EC lead to poorer resolution of the loop structures, although the combination of atomic models with AFM surfaces allows clear interpretation of the extent and flexibility of the loop structures. While XRD remains the premier technique to determine very-high-resolution structures, EC offers a method to determine loop structures unhindered by three-dimensional crystal contacts, and AFM provides information about surface structures and their flexibility under physiological conditions.

Amino Acid Motifs↗

PubMed1998

Atomic force microscopy used for the surface characterization of microcapsule immunoisolation devices.

The surface morphology of the microcapsule used as a bioartificial pancreas was examined by atomic force microscopy (AFM) under ambient conditions in a liquid environment. The standard contact mode was used for imaging. The capsules exhibited different morphologies and surface roughness depending on the composition of the cation solution: namely, the mole ratio of antigelling and gelling cations [Na+]/[Ca2+]. Surface roughness parameters obtained by AFM measurements provide quantitative information on the surface properties of the capsular membrane. In this respect, AFM can be considered a valuable technique complementary to optical microscopy in providing feedback for capsule optimization.

Artificial Organs↗

PubMed2006

Characterization of bioscoured cotton fabrics using FT-IR ATR spectroscopy and microscopy techniques.

The effects of bioscouring were investigated by characterizing the chemical and physical surface changes of cotton fabrics using a purified pectinase enzyme from Bacillus subtilis strain WSHB04-02. Fourier-transform infrared (FT-IR) attenuated total-reflectance (ATR) spectroscopy, scanning electron microscopy (SEM), and atomic force microscopy (AFM) techniques were employed. FT-IR ATR spectroscopy provided a fast and semi-quantitative assessment of the removal of pectins and/or waxes on the cotton surface by comparing the changes in intensity of the carbonyl peak induced by HCl vapor treatment at around 1736 cm(-1). The bioscoured surface could be clearly distinguished from those of untreated and alkali-treated cotton fibers using a combination of SEM and AFM. The images produced using these techniques revealed that the surface morphography and topography of the cotton fibers were shaped by the etching action mode of pectinases during bioscouring. These findings demonstrated that AFM is a useful supplement to SEM in characterizing cotton surfaces.

Bacillus subtilis↗

PubMed1999

AFM imaging of surface adsorbed polymeric 3-alkylpyridinium salts from the marine sponge Reniera sarai.

Bioactive 3-alkylpyridinium polymers (poly-APS) have recently been isolated from the marine sponge Reniera sarai. Previous results have shown that these molecules in aqueous solutions form supramolecular aggregates with an average hydrodynamic radius of 23 +/- 2 nm. To obtain additional evidences about the shape and the dimensions of poly-APS aggregates, we used atomic force microscopy (AFM) operating in tapping mode. The images clearly showed adsorbed aggregates with a lateral dimension of approximately 40 nm and a thickness of the order of approximately 1 nm. The distribution of volumes of the adsorbed aggregates is very similar to the distribution of hydrodynamic radii as obtained from the dynamic light scattering experiments. The volume distribution of these aggregates shows a maximum at 1750 nm3, which corresponds to a sphere with a radius of 7.5 nm.

Adsorption↗

PubMed2006

Characterization of a single molecular QCA cell by Q-control enhanced amplitude modulation atomic force microscopy.

Major technical challenges for reduction of device sizes for computation and memory are the interconnection and power dissipation problems. Molecular quantum-dot cellular automata (QCA) have been proposed as solutions to this problem. Silicon phthalocyanine (SiPc) is a possible candidate for a molecular QCA element. Therefore, it is important to develop an in situ observation technique to visualize individual SiPc molecules. We report here the first image of a single SiPc dimer in air by using quality factor control (Q-control) enhanced amplitude modulation atomic force microscopy (AFM) and an investigation of the interaction forces between the tip and SiPc dimer. The AFM was operated at 0% relative humidity in an ultrapure nitrogen environment either with or without Q-control. Theoretical simulations using the point-mass description of the AFM, demonstrated that Q-enhancement reduced the force exerted from the tip to the sample surface. Our results, consistent with theory, demonstrated that the image force was reduced and a greater height and a larger size were measured. The advantages of this method can be extended to the AFM observation of other "soft" structures, and these results can be useful for a wider community.

Computers, Molecular↗

PubMed2006

AFM and TEM observations of alpha-helix to beta-sheet conformational change occurring on carbon nanotubes.

Bacteriorhodopsin (BR), which is rich in alpha-helical structure, was spread onto water with single-wall carbon nanotubes (SCNTs). After a Langmuir trough was used to apply compressive surface pressure to maintain the alpha-helices monolayer of denatured BR, the composite films comprising alpha-helices and SCNTs were transferred horizontally onto substrates. Atomic force microscopy (AFM) and fluorescence microscopy observation suggested that alpha-helices in contact with SCNTs changed into beta-sheets. High-resolution transmission electron microscopy (HR-TEM) showed 0.54 nm periodicity characteristic of the turn of alpha-helical structure in the SCNTs-free alpha-helix monolayer region and showed the 0.70 nm periodicity of beta-sheet pleated structure in the region where SCNTs were covered with unfolded BR. Unique features of carbon nanotubes that trigger conformational changes of a protein were revealed.

Bacteriorhodopsins↗

PubMed2003

Lubricating properties of the initial salivary pellicle--an AFM study.

The role of saliva in the oral cavity is manifold; an important function is to serve as lubricant between hard (enamel) and soft (mucosal) tissues. Intraoral lubrication is of crucial importance in order to maintain functions such as deglutition, mastication and the faculty of speech. A large number of people suffer from impaired salivary functions, displaying symptoms such as 'dry mouth'. This results in a need for methods to assess the lubricating properties of both native saliva and potential artificial saliva formulations. Here, normal as well as lateral forces, acting between adsorbed salivary films, have been measured for the first time by means of colloidal probe atomic force microscopy (AFM). It was found that the presence of salivary pellicles between hard surfaces reduces the friction coefficient by a factor of 20. This reduction of friction is consistent with the long-range purely repulsive nature of the normal forces acting between the salivary films. The lubricating mechanism is presumably based on a full separation of the sliding surfaces by the salivary films. The friction between salivary films has been investigated at normal loads that cover the clinical jaw closing forces, and it can be concluded that the lubricating properties are maintained within this load interval. The present study indicates the usefulness of colloidal probe AFM, which offers a direct and quantitative measure of lubrication at a molecular level, in the study of biotribological phenomena. In particular, the results obtained here may have implications for the development of saliva substitutes.

Adult↗

PubMed2004

Biofunctionalization of carbon nanotubes for atomic force microscopy imaging.

The study of biological processes relies increasingly on methods for probing structure and function of biochemical machinery (proteins, nucleic acids, and so on) with submolecular resolution. Atomic force microscopy (AFM) has recently emerged as a promising approach for imaging biological structures with resolution approaching the nanometer scale. Two important limitations of AFM in biological imaging are (1) resolution is constrained by probe tip dimensions, and (2) typical probe tips lack chemical specificity to differentiate between functional groups in biological structures. Single-walled carbon nanotubes (SWNTs) offer an intriguing possibility for providing both high resolution and chemical selectivity in AFM imaging, thus overcoming the enumerated limitations. Procedures for generating SWNT tips for AFM will be described. Carboxylic acid functional groups at the SWNT ends can be functionalized using covalent coupling chemistry to attach biological moieties via primary amine groups. Herein, the focus will be on describing methods for attaching biotin to SWNT tips and probing streptavidin on surfaces; importantly, this same coupling chemistry can also be applied to other biomolecules possessing primary amine groups. Underivatized SWNT tips can also provide high-resolution AFM images of DNA. Biofunctionalization of SWNT AFM tips offers great potential to enable high-resolution, chemically selective imaging of biological structures.

Biotin↗

PubMed2003

[The new measurement technics in biology and medicine--atomic force microscopy].

Scanning probe microscopy (SPM) has recently become the most dynamically developing technique employed in research investigations. Atomic force microscopy (AFM) that uses a digitally-controlled scanner and a cantilever system with a measurement tip shaped as a cone with the apex radius of 2-50 nm and the application force of 10(-10) N, as well as a system of recording detectors, has allowed for identification of numerous biological structures in a nanometer scale. The paper presents a review of measurement methods that employ the atomic force microscopy and are being used in medicine and biology, along with major achievements accomplished owing to this technique by various investigators, including the authors themselves.

Animals↗

PubMed1995

AFM and XRD investigation of crystalline vapor-deposited C(60) films.

The morphological and structural properties of C(60) films deposited on quartz substrates by sublimation at 320-500 degrees C under high vacuum have been investigated using atomic force microscopy (AFM) and reflection X-ray diffraction (RXRD). The thickness of the films varied between 0.2 microm and 10 microm. AFM showed that the films consist predominantly of cubic crystals of a few micrometer in size with well-developed (111) and (100) faces. The crystallographic investigation revealed a strongly preferred [111] growth direction which is very sensitive to the deposition rate and substrate temperature. The influence of the experimental parameters on the morphology of the crystals and on the preferred orientation of the films is discussed in view of the AFM and RXRD results.

Journal Article↗

PubMed2004

Generation of amino-terminated surfaces by chemical lithography using atomic force microscopy.

Self-assembled monolayers (SAMs) covered with nitroso end groups were reduced using an atomic force microscope. As the bias voltage become more negative (beyond -4 V), the surface potential of the scanned area become closer to that of the amino-terminated SAM. Following this chemical change, however, no change in topographic features was detected, implying retained stability of the underlying SAM layer. We then released carboxylate-modified polystyrene (PS) spheres into a pH 4 solution containing the sample. Subsequent imaging with atomic force microscopy (AFM) revealed that these PS spheres were only selectively immobilized on the regions that were originally scanned at -6 V to form amino termination. In summary, using AFM set to a specific voltage, we were able to selectively generate micropatterned regions of the SAM with amino termination.

Amines↗

PubMed1997

The atomic force microscope detects ATP-sensitive protein clusters in the plasma membrane of transformed MDCK cells.

Plasma membrane proteins are supposed to form clusters that allow 'functional cross-talk' between individual molecules within nanometre distance. However, such hypothetical protein clusters have not yet been shown directly in native plasma membranes. Therefore, we developed a technique to get access to the inner face of the plasma membrane of cultured transformed kidney (MDCK) cells. The authors applied atomic force microscopy (AFM) to visualize clusters of native proteins protruding from the cytoplasmic membrane surface. We used the K+ channel blocker iberiotoxin (IBTX), a positively charged toxin molecule, that binds with high affinity to plasma membrane potassium channels and to atomically flat mica. Thus, apical plasma membranes could be 'glued' with IBTX to the mica surface with the cytosolic side of the membrane accessible to the scanning AFM tip. The topography of these native inside-out membrane patches was imaged with AFM in electrolyte solution mimicking the cytosol. The plasma membrane could be clearly identified as a lipid bilayer with the characteristic height of 4.9 +/- 0.02 nm. Multiple proteins protruded from the lipid bilayer into the cytosolic space with molecule heights between 1 and 20 nm. Large protrusions were most likely protein clusters. Addition of the proteolytic enzyme pronase to the bath solution led to the disappearance of the proteins within minutes. The metabolic substrate ATP induced a shape-change of the protein clusters and smaller subunits became visible. ADP or the non-hydrolysable ATP analogue, ATP-gamma-S, could not exert similar effects. It is concluded that plasma membrane proteins (and/or membrane associated proteins) form 'functional clusters' in their native environment. The 'physiological' arrangement of the protein molecules within a cluster requires ATP.

Adenosine Triphosphate↗