PubMed Health⌕ Search

Biomedical subjects

M J Miles

Publications and source records attributed to M J Miles.

At least 19 recordsLinked to original sources
PubMed2002

Heterogeneity and persistence length in human ocular mucins.

Atomic force microscopy (AFM) has been used to investigate the heterogeneity and flexibility of human ocular mucins and their subunits. We have paid particular attention, in terms of theory and experiment, to the problem of inducing the polymers to assume equilibrium conformations at a surface. Mucins deposited from a buffer containing Ni(2+) ions adopt extended conformations on mica akin to those observed for DNA under similar conditions. The heterogeneity of the intracellular native mucins is evident from a histogram of contour lengths, reflecting, in part, the diversity of mucin gene products expressed. Reduction of the native mucin with dithiothreitol, thereby breaking the S==S bonds between cysteine residues, causes a marked reduction in polymer length. These results reflect the modes of transport and assembly of newly synthesized mucins in vivo. By modifying the worm-like chain model for applicability to two dimensions, we have confirmed that under the conditions employed mucin adsorbs to mica in an equilibrated conformation. The determined persistence length of the native mucin, 36 nm, is consistent with that of an extended, flexible polymer; such characteristics will influence the properties of the gels formed in vivo.

Air↗
PubMed2001

Internal structure of the starch granule revealed by AFM.

Atomic force microscopy images of sectioned native corn starch granules show evidence of the well-known radial organisation of the starch macromolecules, with the less-ordered hilum region near to the centre. Native granules show blocks 400-500 nm in size that span the growth rings. Lintnerised starch granules, where a mild acid hydrolysis has been used to remove the amorphous and less crystalline parts of the granule, clearly show smaller 'blocklets' within the rings approximately 10-30 nm in size. This level of organisation within the growth rings corresponds to the blocklet or superhelix structures that have been proposed in the literature for the association or clustering of amylopectin helices. Mechanical property imaging techniques have provided enhanced contrast to view this morphology, and shown the deformability of the starch structure under contact mode imaging conditions.

Amylopectin↗
PubMed2001

High-Q dynamic force microscopy in liquid and its application to living cells.

We present a new dynamic force microscopy technique for imaging in liquids in the piconewton regime. The low quality factor (Q) of the cantilever is increased up to three orders of magnitude by the implementation of a positive feedback control. The technique also includes a phase-locked loop unit to track the resonance of the cantilever. Experiments and computer simulations indicate that the tip-sample forces are below 100 pN, about two orders of magnitude lower than in conventional tapping mode atomic force microscopy. Furthermore, the spectroscopic ability is greatly enhanced. Either the phase shift or the resonant frequency shows a high sensitivity to variations in either the energy dissipation or conservative interactions between the tip and the sample, respectively. The potential of this technique is demonstrated by imaging living cells.

Animals↗
PubMed2001

Chemical sensors and biosensors in liquid environment based on microcantilevers with amplified quality factor.

A new technique is presented for bio/chemical sensors, based on microcantilevers, for detection in liquid environment. The low quality factor of the cantilever in liquid is increased up to three orders of magnitude by using Q-control. This enables AC detection that is immune to the long-term drift of the DC cantilever response in liquids, and to temperature variations. This technique has been applied for the detection of ethanol in aqueous solution by using the microbalance method, and for antibody/antigen recognition by the surface stress method. The results show the feasibility and very high sensitivity of these novel devices.

Antibodies↗
PubMed2001

Modeling of cylindrically tapered cantilevers for transverse dynamic force microscopy (TDFM).

In transverse dynamic force microscopy a cylindrically tapered cantilever is mounted perpendicularly to the sample surface and set into transversal oscillation. The dynamics of the cantilever has been studied using the continuum mechanical model with discrete element analysis. A viscoelastic model has been used to describe the tip-sample interaction. In this way an in-phase and an out-of-phase component of the force has been extracted from the experimental data. Two different techniques, involving two experimental setups and two corresponding data analysis routines, have been developed to calculate the two components of the force at different tip-sample separations. In one case the change in resonant frequency and corresponding oscillation amplitude is measured whereas in the second case the usual way of recording amplitude and phase signal at a fixed driving frequency is applied. The results from these two methods are shown to be completely consistent and produce almost identical force curves.

Journal Article↗
PubMed2001

Exploring the molecular adhesion of ocular mucins.

Mucins have been ascribed both pro- and anti-adhesive functions. To clarify how both functions can be embodied in the same molecule we studied the interaction of human ocular mucins with mica and with mucins deposited on mica. Adhesion energy and forces of interaction were evaluated as a function of speed of approach, dwell time at maximum extension, and presence of divalent cations in the imaging buffer. Mucins were tethered to an AFM gold-coated tip. Repeated cycles of approach and retract to mica revealed a large number of adhesions in each cycle. Adhesion energy (0.2-48 aJ) and detachment forces (0.1-4 nN) increased with the addition of Ni(II) ions, and with lengthening dwell time. Speed of approach made little difference to the interactions. Most detachments occurred less than 40 nm from the surface. Inter-detachment distances reflected the major periodicities of the mica basal plane. Short distances of interaction, magnitude of detachment forces, and imaging of mucins on SAM all suggest deformable compact mucin aggregates on the AFM tip. Inter-detachment distances suggest a large degree of interpenetration between neighboring molecules. Tip-tethered mucins did not adhere to mucins deposited on mica. This phenomenon is analogous with the nonadherence of the mucin gels on lids and on cornea during blinking.

Aluminum Silicates↗
PubMed2000

New insight into cellulose structure by atomic force microscopy shows the i(alpha) crystal phase at near-atomic resolution.

The organization of the surface of cellulose is important in cell structure, as well as in industrial processing and modification. Using atomic force microscopy, we show that the I(alpha) phase of native cellulose first proposed in 1984 and subsequently characterized by a triclinic unit cell exists over large areas of the surface of microcrystals from Valonia, one of the most highly crystalline celluloses. There is startling agreement between the observed structure and crystal models, and it is possible to identify the specific crystal face being imaged. The near-atomic resolution images also offer an insight into structural reconstructions at the surface compared to the interior. We are able to assign features in the images to particular side groups attached to the glucose ring and find indications of subtle modifications of the position of surface hydroxyls due to changes in hydrogen bonding.

Carbohydrate Conformation↗
PubMed1999

Small angle X-ray scattering of wheat seed-storage proteins: alpha-, gamma- and omega-gliadins and the high molecular weight (HMW) subunits of glutenin.

Small angle X-ray scattering in solution was performed on seed-storage proteins from wheat. Three different groups of gliadins (alpha-, gamma- and omega-) and a high molecular weight (HMW) subunit of glutenin (1Bx20) were studied to determine molecular size parameters. All the gliadins could be modelled as prolate ellipsoids with extended conformations. The HMW subunit existed as a highly extended rod-like particle in solution with a length of about 69 nm and a diameter of about 6.4 nm. Specific aggregation effects were observed which may reflect mechanisms of self-assembly that contribute to the unique viscoelastic properties of wheat dough.

Gliadin↗
PubMed1999

Atomic force microscopy of the submolecular architecture of hydrated ocular mucins.

High-resolution atomic force microscopy has been applied to the imaging of intact human ocular mucins in a near-physiological buffer. The mucins displayed a range of lengths from several hundred nanometers to several microns. By varying the ionic composition of the imaging environment, it was possible to image molecules rigidly fixed to the substrate and the motion of single molecules across the substrate. From static molecular images, high-resolution line profiles show a variation of up to +/-0.75 nm in thickness along the molecule. This variation is localized in regions of several tens of nanometers. It is interpreted in terms of the varying glycosylation along the mucin and is consistent with the known size of oligosaccharides in ocular mucins. The dynamic images indicate the possibility of following mucin interactions in situ.

Biopolymers↗
PubMed1999

Identification of microphases in mixed alpha- and omega-gliadin protein films investigated by atomic force microscopy.

Pure and mixed films of alpha- and omega-gliadins were studied by tapping mode atomic force microscopy (AFM). The technique was sensitive to the chemistry of the surface properties of the films, allowing imaging of the mixed gliadin phases at different ratios. In addition to the study of the phases at the micrometer level, higher resolution images allowed visualization of the protein films at the molecular level. These studies may have relevance to the formation of phases in developing protein bodies in grain, where gliadins and glutenins are deposited together. It has been assumed that the protein bodies consist of a random network of proteins; these studies indicate that microphases could be present in protein bodies. The technique provides novel methods for studying mixed biopolymer systems.

Gliadin↗
PubMed1996

Direct observation of protein secondary structure in gas vesicles by atomic force microscopy.

The protein that forms the gas vesicle in the cyanobacterium Anabaena flos-aquae has been imaged by atomic force microscopy (AFM) under liquid at room temperature. The protein constitutes "ribs" which, stacked together, form the hollow cylindrical tube and conical end caps of the gas vesicle. By operating the microscope in deflection mode, it has been possible to achieve sub-nanometer resolution of the rib structure. The lateral spacing of the ribs was found to be 4.6 +/- 0.1 nm. At higher resolution the ribs are observed to consist of pairs of lines at an angle of approximately 55 degrees to the rib axis, with a repeat distance between each line of 0.57 +/- 0.05 nm along the rib axis. These observed dimensions and periodicities are consistent with those determined from previous x-ray diffraction studies, indicating that the protein is arranged in beta-chains crossing the rib at an angle of 55 degrees to the rib axis. The AFM results confirm the x-ray data and represent the first direct images of a beta-sheet protein secondary structure using this technique. The orientation of the GvpA protein component of the structure and the extent of this protein across the ribs have been established for the first time.

Anabaena↗
PubMed1995

Atomic force microscopy of plant chromosomes.

Atomic force microscopy has been used to image plant chromosomes from standard preparations without staining or coating. This has enabled the collection of high-resolution three-dimensional data on surface structure. The technique has been further applied to the imaging of C-banded chromosomes revealing structural changes resulting from the banding treatment. The bands were observed as localized areas of high relief.

Chromosome Banding↗
PubMed1995

Atomic force microscopy of the myosin molecule.

Atomic force microscopy (AFM) has been used to study the structure of rabbit skeletal muscle myosin deposited onto a mica substrate from glycerol solution. Images of the myosin molecule have been obtained using contact mode AFM with the sample immersed in propanol. The molecules have two heads at one end of a long tail and have an appearance similar to those prepared by glycerol deposition techniques for electron microscopy, except that the separation of the two heads is not so well defined. The average length of the tail (155 +/- 5 nm) agrees well with previous studies. Bends in the myosin tail have been observed at locations similar to those observed in the electron microscope. By raising the applied force, it has been possible locally to separate the two strands of the alpha-helical coiled-coil tail. We conclude that the glycerol-mica technique is a useful tool for the preparation of fibrous proteins for examination by scanning probe microscopy.

Aluminum Silicates↗
PubMed1994

Application of scanning force microscopy to chromosome analysis.

The application of scanning force microscopy (SFM) to the imaging and analysis of chromosomes is described. This relatively new microscopical technique has been used to provide high-resolution, three-dimensional images of uncoated and unstained human chromosomes in which surface features less than 50 nm have been resolved. Comparison of SFM images with light microscopy data has permitted identification of specific chromosomes, and images of a chromosome showing a cytogenetic abnormality are presented. These preliminary results demonstrate that this technique has potential applications in the imaging and analysis of chromosome structure.

Chromosomes, Human↗
PubMed1992

Visualization of the algal polysaccharide carrageenan by scanning tunnelling microscopy.

Scanning tunnelling microscopy has been used to obtain images in the constant-current mode in air and moist conditions at molecular resolution for the kappa- and iota-carrageenan algal polysaccharides. The molecules were deposited from an aqueous solution onto a graphite substrate. The samples formed aligned nematic-like arrays and were also found as individual molecules. The molecular dimensions of width, height and repeat distance along the molecule were found to be close to the values previously determined by X-ray diffraction. The results support a molecular model based on a double-helix structure for carrageenan.

Carbohydrate Sequence↗