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At least 19 recordsLinked to original sources
PubMed1998

Preparation of basal cell membranes for scanning probe microscopy.

Scanning probe microscopy has the potential for investigating membranes in a physiological environment. We prepared with a lysis-squirting protocol basal cell membranes, that are suitable for scanning probe microscopy. Investigations using atomic force microscopy under liquid revealed cellular filaments which correlated perfectly with fluorescently stained actin filaments. Globular structures with a diameter as little as 10 nm could be resolved by stripping cytoplasmic components from the membranes. Therefore, cytoplasmic sides of supported basal cell membranes prove useful to gain high resolution with scanning probe microscopy in studies of plasma membrane associated structures and processes under buffer solution.

Animals↗
PubMed1997

Adsorption of biological molecules to a solid support for scanning probe microscopy.

Scanning probe microscopes are now established tools to study the surface structure of biological macromolecules under physiological conditions. Sample preparation methods for this microscopy all have the objective to attach the specimen firmly to a support. Here we analyse the commonly used method of adsorbing biological specimens to freshly cleaved mica. This is facilitated by adjusting the electrolyte concentration and the pH of the buffer solution. Native macromolecular systems absorbed to mica in this way can be reproducibly imaged at submolecular resolution.

Adsorption↗
PubMed2000

Ultra Flat Gold Surfaces for Use in Chemical Force Microscopy: Scanning Probe Microscopy Studies of the Effect of Preparation Regime on Surface Morphology.

The preparation of ultra flat gold surfaces for use in chemical force microscopy (CFM) has been studied. The surfaces were studied in terms of substrate effects by comparing mica, Si (110) wafer and glass slides. The effect of different annealing regimes was also investigated. Measurements on these surfaces were made by both atomic force microscopy (AFM) (in contact and tapping mode) and by scanning tunneling microscopy (STM). The films contain different morphologies with respect to grain size and topography. Calculations of surface roughness present values less than 2.5 nm for all surfaces studied, making the choice of the "flattest" surface difficult if based on criteria of surface roughness alone. Additionally, it is shown that different acquisition parameters can produce dissimilar images that have stability and reproducibility.

Journal Article↗
PubMed1993

BCG cell imaging using scanning probe microscopy.

Scanning tunneling microscopy (STM) and atomic force microscopy (AFM) were used to obtain images of the surface of whole, intact BCG (bacille Calmette Guerin, a mycobacterium) cells in air and under solution by immobilizing the cells onto glass slides (AFM only) or highly oriented pyrolytic graphite. The technique used for AFM imaging involved depositing a submonolayer of cells under a centrifugal force followed by fixation/dehydration using polar organic solvents. AFM images agree well with images from light and electron microscopy and showed large numbers of BCG cells in their distinctive cord arrangement. The AFM also proved useful for identifying extracellular microgranules which cannot be seen with light microscopy. For STM imaging, the hydrophobicity of BCG enabled strong adhesion from aqueous solution onto graphite. STM images of BCG could only be obtained while scanning in aqueous solution, and the cells showed a large variation in contrast when different samples were imaged. The STM provided greater detail of surface features than the AFM and was able to produce images of periodic layers corroborating observations made by transmission electron microscopy.

Microscopy↗
PubMed1995

Scanning probe microscopy in microbiology.

Scanning probe microscopy (SPM) is emerging as an important alternative to electron microscopy as a technique for analyzing submicron details on biological surfaces. Microbiological specimens such as viruses, bacteriophages, and ordered bacterial surface layers and membranes have played an important role in the development of scanning tunnelling microscopy (STM) and atomic force microscopy (AFM) in cellular and molecular biology. Early STM studies involving metal-coated bacteriophage T4 polyheads, Methanospirillum hungatei, and Deinococcus radiodurans HPI layer clearly demonstrated that resolution was comparable to TEM on similarly prepared specimens and only limited by metal graininess. However, except for thin films or layers, novel biological information has been difficult to obtain since imaging native surfaces of such biomaterials as proteins or nucleic acids by STM proved to be unreliable. With the development of atomic force microscopes, which allow imaging of similar native structures, SPM applications have widened to include straightforward surface structure analysis, analysis of surface elastic and inelastic properties, bonding force measurements between molecules, and micro-manipulations of such individual molecules as DNA. AFM images have progressed from relatively crude representations of specimen topography to nanometer scale representations of native hydrated surfaces. It appears from the study of microbiological specimens that direct visualization of dynamic molecular events or processes may soon become a reality.

Bacteria↗
PubMed1998

Atomic force microscopy and other scanning probe microscopies.

The highlight of the past year is the unfolding and refolding of the muscle protein titin in the atomic force microscope. A related highlight in the intersection between experiment and theory is a recent review of the effects of molecular forces on biochemical kinetics. Other advances in scanning probe microscopy include entropic brushes, molecular sandwiches and applications of atomic force microscopy to gene therapy.

Annexin A5↗
PubMed1992

Immobilization of DNA for scanning probe microscopy.

Reproducible scanning tunneling microscope and atomic force microscope images of entire molecules of uncoated plasmid DNA chemically bound to surfaces are presented. The chemically mediated immobilization of DNA to surfaces and subsequent scanning tunneling microscope imaging of DNA molecules demonstrate that the problem of molecular instability to forces exerted by the probe tip, inherent with scanning probe microscopes, can be prevented.

DNA, Bacterial↗
PubMed1997

Scanning probe microscopy.

During the past year, scanning probe microscopy, especially atomic force microscopy (AFM), has taken root in the biological sciences community, as is evident from the large number of publications and from the variety of specialized journals in which these papers appear. Furthermore, there is a strong indication that the technique is evolving from a qualitative imaging tool to a probe of the critical dimensions and properties of biomolecules and living cells. The next stage of the evolution involves the development of microinstruments for process control and sensing applications. Recent advances have been reported in AFM instrumentation and method. For example, the tapping mode of operation is becoming the method of choice to image biological molecules; work to extend tapping-mode operation in liquids has been reported. Biological molecules can also be imaged at low temperature in a cryo-AFM with improved resolution. The measurement of recognition forces between individual molecules continues to attract much attention and has spawned new concepts for ultra-sensitive biosensors. The AFM is being used increasingly for property measurements such as determining the viscoelastic properties of biological molecules. Finally, structural studies using the AFM abound. Some specific highlights include the mapping of DNA using restriction enzymes, imaging during DNA transcription and determining the mode of drug binding to DNA.

Cell Membrane↗
PubMed2000

Structural differences in unbleached and mildly-bleached synthetic tyrosine-derived melanins identified by scanning probe microscopies.

Using scanning tunneling microscopy (STM), we have imaged two types of mildly-bleached, synthetic tyrosine-derived melanins for comparison with the unbleached melanin from which they were prepared. These mildly-bleached melanins were generated by mild oxidation of the unbleached melanin, using either basic hydrogen peroxide or air/light. The unbleached melanin, and two mildly-bleached melanins, were independently deposited from very dilute tetrahydrofuran (THF) solutions onto highly oriented pyrolytic graphite (HOPG) substrate for STM imaging. Lateral dimensions (23 A, average of two directions) of structures from each of the three samples showed no differences. However, structures from both mildly-bleached melanins showed similar dramatic decreases (from approximately 15 A to approximately 5 A) in their STM-measured apparent heights, compared with structures from the unbleached melanin sample. These STM observations are compatible with structural models for unbleached and mildly-bleached melanins, incorporating a three-dimensional structure for unbleached melanin composed of multi-layered, pi-pi-stacked, carboxylic and amino variants of polyaromatic polymeric sheets. The STM-observed decrease in apparent heights after mild oxidation, which we associate with a change in stack height, has been confirmed by experiments using tapping mode atomic force microscopy (TM-AFM) for the unbleached and mildly-hydrogen-peroXide-bleached melanins (from approximately 14 A to approximately 6 A). In these TM-AFM experiments, the melanins were deposited directly onto magnesium cation-treated glass substrates in contact with methanolic solutions of each of the melanins. We interpret our mild-bleaching results as an oxidative conversion of the multi-layered, stacked sheets of mainly carboxylic and amino variants of polyquinhydrone-like moieties, to largely de-stacked, mildly-bleached melanin sheets. These oxidized and, hence, electron-deficient sheets should not readily form multi-layered, pi-pi interacting stacks, but instead appear to be either single-layer polyquinone sheets or, at most, double-layer polyquinhydrone sheets. The effects of such de-stacking on in vivo melanin photoprotection, and structural similarities between melanin derived from natural sources and the synthetic melanin samples used in this work are discussed.

Absorption↗
PubMed1997

Scanning probe microscopy for the characterization of biomaterials and biological interactions.

The scanning probe microscopies provide a unique view of biological and biomedical systems at a nanoscale appropriate to appreciate molecular events. The advent of these methods has brought the ability to acquire quantitative information at the molecular level. Given the proliferation of microscopes and associated methods, the probability for important discoveries is high. If tempered with an appreciation for the potential for artifacts, the SPMs may revolutionize our view of biological systems and biomaterials interactions with those systems.

Animals↗
PubMed2000

The importance of molecular structure and conformation: learning with scanning probe microscopy.

Molecular structure holds a key to understanding Nature's intricate design mechanisms and blueprints. If we can understand her blueprints and basic materials, perhaps we can begin to mimic her beautiful products more cost effectively and with less detrimental environmental consequences. Higher resolution instrumentation has allowed us to study single molecules. Indeed, many stellar contributions to the field have come forth in the last couple of years. We can measure the forces required to unravel individual domains of biological molecules such as titin or DNA to a few picoNewtons resolution. This review will attempt to provide a general overview of the field of single molecule analysis using scanning force microscopy.

Adhesives↗
PubMed1996

Progress in the application of scanning probe microscopy to biology.

Several key developments have occurred recently in the application of scanning probe microscopy to biology. These include the use of 'tapping-mode' atomic force microscopy both for the high-resolution imaging of biomolecules in liquids and for monitoring in situ biocatalysis, the use of atomic force microscopy as a force transducer to measure individual biological interactions, and the development of hybrid techniques such as scanning tunnelling microscopy coupled to confocal scanning laser microscopy.

Journal Article↗
PubMed1997

Data analysis using the Internet: the World Wide Web scanning probe microscopy data analysis system.

The first interactive world-wide web-based image analysis system is presented (http://pharm6.pharm.nottingham.ac.uk/processing/main. html). The system, currently tailored to scanning probe microscopy image data, has been developed to permit the use of software algorithms developed within our laboratory by researchers throughout the world. The implementation and functionality of the scanning probe microscopy server is described. Feedback from users of the facility has demonstrated its value within the research community, and highlighted key operational issues which are to be addressed. A future role of Internet-based data processing software is also discussed.

Algorithms↗
PubMedDate not supplied

[Scanning probe microscopy and medico-biological nanotechnology: history and prospects].

Literature data on the use of scanning probe microscopy for biological samples studying are presented. As an illustration, some results of the authors' investigations of the shark olfactory receptor surface, human lipoprotein of low density and human erythrocyte are given. Perspectives of the medico-biological development of nanotechnologies are evaluated.

Animals↗
PubMed2000

Scanned probe microscopy of electronic transport in carbon nanotubes.

We use electrostatic force microscopy and scanned gate microscopy to probe the conducting properties of carbon nanotubes at room temperature. Multiwalled carbon nanotubes are shown to be diffusive conductors, while metallic single-walled carbon nanotubes are ballistic conductors over micron lengths. Semiconducting single-walled carbon nanotubes are shown to have a series of large barriers to conduction along their length. These measurements are also used to probe the contact resistance and locate breaks in carbon nanotube circuits.

Journal Article↗