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At least 973 records · Page 54Linked to original sources
PubMed2003

Electrostatic nanolithography in polymers using atomic force microscopy.

The past decade has witnessed an explosion of techniques used to pattern polymers on the nano (1-100 nm) and submicrometre (100-1,000 nm) scale, driven by the extensive versatility of polymers for diverse applications, such as molecular electronics, data storage, optoelectronics, displays, sacrificial templates and all forms of sensors. Conceptually, most of the patterning techniques, including microcontact printing (soft lithography), photolithography, electron-beam lithography, block-copolymer templating and dip-pen lithography, are based on the spatially selective removal or formation/deposition of polymer. Here, we demonstrate an alternative and novel lithography technique--electrostatic nanolithography using atomic force microscopy--that generates features by mass transport of polymer within an initially uniform, planar film without chemical crosslinking, substantial polymer degradation or ablation. The combination of localized softening of attolitres (10(2)-10(5) nm3) of polymer by Joule heating, extremely non-uniform electric field gradients to polarize and manipulate the soften polymer, and single-step process methodology using conventional atomic force microscopy (AFM) equipment, establishes a new paradigm for polymer nanolithography, allowing rapid (of the order of milliseconds) creation of raised (or depressed) features without external heating of a polymer film or AFM tip-film contact.

Microscopy, Atomic Force↗
PubMed2003

Atomic force microscopy investigation of human immunodeficiency virus (HIV) and HIV-infected lymphocytes.

Isolated human immunodeficiency virus (HIV) and HIV-infected human lymphocytes in culture have been imaged for the first time by atomic force microscopy (AFM). Purified virus particles spread on glass substrates are roughly spherical, reasonably uniform, though pleomorphic in appearance, and have diameters of about 120 nm. Similar particles are also seen on infected cell surfaces, but morphologies and sizes are considerably more varied, possibly a reflection of the budding process. The surfaces of HIV particles exhibit "tufts" of protein, presumably gp120, which do not physically resemble spikes. The protein tufts, which number about 100 per particle, have average diameters of about 200 A, but with a large variance. They likely consist of arbitrary associations of small numbers of gp120 monomers on the surface. In examining several hundred virus particles, we found no evidence that the gp120 monomers form threefold symmetric trimers. Although >95% of HIV-infected H9 lymphocytic cells were producing HIV antigens by immunofluorescent assay, most lymphocytes displayed few or no virus on their surfaces, while others were almost covered by a hundred or more viruses, suggesting a dependence on cell cycle or physiology. HIV-infected cells treated with a viral protease inhibitor and their progeny viruses were also imaged by AFM and were indistinguishable from untreated virions. Isolated HIV virions were disrupted by exposure to mild neutral detergents (Tween 20 and CHAPS) at concentrations from 0.25 to 2.0%. Among the products observed were intact virions, the remnants of completely degraded virions, and partially disrupted particles that lacked sectors of surface proteins as well as virions that were split or broken open to reveal their empty interiors. Capsids containing nucleic acid were not seen, suggesting that the capsids were even more fragile than the envelope and were totally degraded and lost. From these images, a good estimate of the thickness of the envelope protein-membrane-matrix protein outer shell of the virion was obtained. Treatment with even low concentrations (<0.1%) of sodium dodecyl sulfate completely destroyed all virions but produced many interesting products, including aggregates of viral proteins with strands of nucleic acid.

Cell Line↗
PubMed2002

Potassium-selective atomic force microscopy on ion-releasing substrates and living cells.

A new method for simultaneous mapping of cell topography and ion fluxes was developed. A highly sensitive ion sensor system was generated by coating atomic force microscopy tips with a PVC layer containing valinomycin, an ionophore for potassium. The activity of specific ions was traced on artificial ion-releasing PVC substrates. A boundary potential was generated owing to the selective exchange of a specific ion between coated tip and ion-releasing substrate. The boundary potential was detectable as a force induced by ion-selective electrostatic interactions. The selectivity coefficient of valinomycin for potassium against sodium (K(K,Na)f) was -2.5 +/- 0.5. Potassium efflux was measured on living MDCK-F1 cells expressing BK(Ca) channels. We could demonstrate localized areas of high potassium concentrations at the cell surface. The potassium efflux could be reversibly inhibited by thapsigargin, which is known to inhibit the efflux of potassium from BK(Ca) channels by suppression of calcium ATPase.

Biosensing Techniques↗
PubMedDate not supplied

Fine-stranded and particulate aggregates of heat-denatured whey proteins visualized by atomic force microscopy.

beta-Lactoglobulin and whey protein isolate (WPI) were heated in aqueous solutions at pH 2 and 7 at 80 degrees C, spread onto freshly cleaved mica surfaces, and visualized under butanol using atomic force microscopy. Fine-stranded aggregates were formed at pH 2, the diameter of strands being ca. 4 nm for beta-lactoglobulin and 10 nm for WPI. At pH 7, aggregates were composed of ellipsoidal particles, regardless of the concentration of added NaCl. This observation supports the previously proposed two-step aggregation model at neutral pH (Aymard, P.; Gimel, J. C.; Nicolai, T.; Durand, D. J. Chim. Phys. 1996, 93, 987-997), consisting of the formation of primary globular particles and the subsequent aggregation of those primary particles. The AFM provides the first direct evidence for the anisotropic shape of these primary particles. The heights of primary particles increased from ca. 11 to 27 nm with increasing concentrations of added NaCl from 0 to 0.3 M in the case of WPI. The rate of aggregation was also accelerated with increasing NaCl concentrations, which appeared to induce transitions in gel networks from fine-stranded toward particulate networks. The present study provides structural information essential for understanding the diverse physical properties of heat-induced whey protein gels.

Hot Temperature↗
PubMed2002

Hierarchical assembly and the onset of banding in fibrous long spacing collagen revealed by atomic force microscopy.

The mechanism of formation of fibrillar collagen with a banding periodicity much greater than the 67 nm of native collagen, i.e. the so-called fibrous long spacing (FLS) collagen, has been speculated upon, but has not been previously studied experimentally from a detailed structural perspective. In vitro, such fibrils, with banding periodicity of approximately 270 nm, may be produced by dialysis of an acidic solution of type I collagen and alpha(1)-acid glycoprotein against deionized water. FLS collagen assembly was investigated by visualization of assembly intermediates that were formed during the course of dialysis using atomic force microscopy. Below pH 4, thin, curly nonbanded fibrils were formed. When the dialysis solution reached approximately pH 4, thin, filamentous structures that showed protrusions spaced at approximately 270 nm were seen. As the pH increased, these protofibrils appeared to associate loosely into larger fibrils with clear approximately 270 nm banding which increased in diameter and compactness, such that by approximately pH 4.6, mature FLS collagen fibrils begin to be observed with increasing frequency. These results suggest that there are aspects of a stepwise process in the formation of FLS collagen, and that the banding pattern arises quite early and very specifically in this process. It is proposed that typical 4D-period staggered microfibril subunits assemble laterally with minimal stagger between adjacent fibrils. alpha(1)-Acid glycoprotein presumably promotes this otherwise abnormal lateral assembly over native-type self-assembly. Cocoon-like fibrils, which are hundreds of nanometers in diameter and 10-20 microm in length, were found to coexist with mature FLS fibrils.

Animals↗
PubMed1998

Visualization of PEO-PBLA-pyrene polymeric micelles by atomic force microscopy.

PURPOSE: To directly visualize and evaluate the aqueous block copolymeric micelles, poly(ethylene oxide)-poly(beta-benzyl L-aspartate) (PEO-PBLA) chemically conjugated with pyrene fluorescence molecule, by nanotechnology of atomic force microscopy (AFM). METHODS: The block copolymers' PEO-PBLA-Pyrene was first synthesized by reacting with pyrene sulfonyl chloride and PEO-PBLA in tetrahydrofuran (THF) solution and were identified by GPC reflect index, UV and fluorescence detectors. The characterization of physical and chemical properties of PEO-PBLA-Pyrene polymeric micellar solution were examined by the dynamic light scattering (DLS) and critical micelles concentrations (CMC). In addition, the nanotechnology of AFM was used to directly visualize the size and shape of nanopolymeric micelles. RESULTS: The pyrene fluorescence molecule were successfully conjugated at the amino group of the end of PBLA chain by GPC with three different detectors. The size of the aqueous PEO-PBLA-Pyrene polymeric micelles was detected around 57 nm with unimodal distribution by DLS measurement. As a result of this finding, the CMC test was also found out that the fluorescence intensity was increasing around 0.01 approximately 0.05 mg/ml. Using AFM evaluation of polymeric micellar solution, the morphology of aqueous PEO-PBLA-Pyrene polymeric micelles was observed on round shape and with the narrow dispersity of size range 50 approximately 80 nm. CONCLUSIONS: The presence of PEO-PBLA copolymers with pyrene in an aqueous system formed in a spherical and nano range of polymeric micelles.

Micelles↗
PubMed2001

Shape and size of isolated bone mineralites measured using atomic force microscopy.

The inorganic phase of bone is comprised primarily of very small mineralites. The size and shape of these mineralites play fundamental roles in maintaining ionic homeostasis and in the biomechanical function of bone. Using atomic force microscopy, we have obtained direct three-dimensional visual evidence of the size and shape of native protein-free mineralites isolated from mature bovine bone. Approximately 98% of the mineralites are less than 2 nm thick displaying a plate-like habit. Distributions of both thickness and width show single peaks. The distribution of lengths may be multimodal with distinct peaks separated by approximately 6 nm. Application of our results is expected to be of use in the design of novel orthopaedic biomaterials. In addition, they provide more accurate inputs to molecular-scale models aimed at predicting the physiological and mechanical behavior of bone.

Animals↗
PubMed1994

Slow cellular dynamics in MDCK and R5 cells monitored by time-lapse atomic force microscopy.

We have examined dynamic events that occur on a time scale of minutes in an epithelial monolayer of Madine-Darby Canine Kidney (MDCK) cells and in ras-transformed MDCK cells by atomic force microscopy (AFM). Cells were imaged under physiological conditions, and time-lapse movies representing approximately 60 s real time per frame were assembled. In normal MDCK cells, two types of protrusions in the apical plasma membrane exhibit dynamic behavior. First, smooth bulges formed transiently over the time scale of minutes to tens of minutes. Second, spike-like protrusions appear initially as bulges, extend well above the apical surface and, finally, seem to detach. R5, an oncogenic transformant derived from MDCK cells, grows very flat on glass. During AFM imaging, these cells sometimes round up and detach from the substrate. In light microscopic observations of parallel preparations, cells rarely detach, suggesting that this is an active response of these cells to irritation by the AFM tip. R5 cells often extend processes that are supported by actin stress fibers. During imaging with the AFM, these processes withdraw at a rate of 1-5 microns/min, similar to that observed by light microscopy. During the withdrawal, movement of the stress fibers can be clearly seen. In the flat periphery of these cells, the transport of intracellular particles along cytoskeletal elements was seen. In addition, we have observed two types of wave-like movements through the cell, which appear to be an organized rearrangement of cytoplasm. One type of wave moves radially out from center of the cell while the other moves circularly along the cell periphery.

Animals↗
PubMed2003

Investigations into the polymorphism of rat tail tendon fibrils using atomic force microscopy.

Collagen type I displays a typical banding periodicity of 67 nm when visualized by atomic force or transmission electron microscopy imaging. We have investigated collagen fibers extracted from rat tail tendons using atomic force microscopy, under different ionic and pH conditions. The majority of the fibers reproduce the typical wavy structure with 67 nm spacing and a height difference between the peak and the grooves of at least 5 nm. However, we were also able to individuate two other banding patterns with 23+/-2 nm and 210+/-15 nm periodicities. The small pattern showed height differences of about 2 nm, whereas the large pattern seems to be a superposition of the 67 nm periodicity showing height differences of about 20 nm. Furthermore, we could show that at pH values of 3 and below the fibril structure gets dissolved whereas high concentrations of NaCl and CaCl(2) could prevent this effect.

Animals↗
PubMed2006

Electronic properties of a single-walled carbon nanotube/150mer-porphyrin system measured by point-contact current imaging atomic force microscopy.

The electronic properties of a single-walled carbon nanotube/150mer of porphyrin polymer wire system were investigated. Current-voltage (I-V) curves were measured simultaneously along with topographic observations using point-contact current imaging atomic force microscopy. Symmetric I-V curves were obtained at bare single-walled carbon nanotubes but characteristic asymmetrical rectifying behavior was found at the single-walled carbon nanotube/150mer-porphyrin junctions. This finding is of key importance for the development of new nanoscale molecular electronic devices.

Adsorption↗
PubMed2004

Effect of different anti-Abeta antibodies on Abeta fibrillogenesis as assessed by atomic force microscopy.

Extensive data suggest that the conversion of the amyloid-beta (Abeta) peptide from soluble to insoluble forms is a key factor in the pathogenesis of Alzheimer's disease (AD). In recent years, atomic force microscopy (AFM) has provided useful insights into the physicochemical processes involving Abeta morphology, and it can now be used to explore factors that either inhibit or promote fibrillogenesis. We used ex situ AFM to explore the impact of anti-Abeta antibodies directed against different domains of Abeta on fibril formation. For the AFM studies, two monoclonal antibodies (m3D6 and m266.2) were incubated in solution with Abeta(1-42) with a molar ratio of 1:10 (antibody to Abeta) over several days. Fibril formation was analyzed quantitatively by determining the number of fibrils per microm(2) and by aggregate size analysis. m3D6, which is directed against an N-terminal domain of Abeta (amino acid residues 1-5) slowed down fibril formation. However, m266.2, which is directed against the central domain of Abeta (amino acid residues 13-28) appeared to completely prevent the formation of fibrils over the course of the experiment. Inhibition of fibril formation by both antibodies was also confirmed by thioflavin-T (ThT) fluorescence experiments carried out with Abeta(1-40) incubated for five days. However, unlike AFM results, ThT did not differentiate between the samples incubated with m3D6 versus m266.2. These results indicate that AFM can be not only reliably used to study the effect of different molecules on Abeta aggregation, but that it can provide additional information such as the role of epitope specificity of antibodies as potential inhibitors of fibril formation.

Alzheimer Disease↗
PubMed1998

An integrated approach to the study of living cells by atomic force microscopy.

We describe a technique for studying living cells with the atomic force microscope (AFM) in tapping mode using a thermostated, controlled-environment culture system. We also describe the integration of the AFM with bright field, epifluorescence and surface interference microscopy, achieving the highest level of integration for the AFM thus far described. We succeeded in the continuous, longterm imaging of relatively flat but very fragile cytoplasmic regions of COS cells at a lateral resolution of about 70 nm and a vertical resolution of about 3 nm. In addition, we demonstrate the applicability of our technology for continuous force volume imaging of cultured vertebrate cells. The hybrid instrument we describe can be used to collect simultaneously a diverse variety of physical, chemical and morphological data on living vertebrate cells. The integration of light microscopy with AFM and steady-state culture methods for vertebrate cells represents a new approach for studies in cell biology and physiology.

Animals↗
PubMed2007

Evaluation of surface characteristics of dental composites using profilometry, scanning electron, atomic force microscopy and gloss-meter.

The aim of this in vitro investigation was to compare various roughness and topography measurement methods to characterize the surface quality in several types of resin composites. The initial surface quality of several resin composites was compared. The materials evaluated were of three categories: i) hybrid: TPH Spectrum; ii) reinforced microfill: Micronew and iii) microhybrid: Synergy Duo, Esthet-X, Point.4 and Palfique Estelite. Three Groups of identical disk-shaped specimens (10 x 1.5 mm) were prepared from each material (n = 6) and polished with Soflex discs. Macro-roughness (Ra) was measured with Group 1 by 2-D profilometry. Atomic Force Microscopy (AFM) gave 3-D images and micro-roughness (Ra) of Group 2. Surface optical gloss at 60 degrees was determined for Group 3. Specimens of each material were also studied by scanning electron microscopy. Macro-Ra values (microm) ranged from 0.30 to 0.56. Micro-Ra values ranged from 0.03 to 0.14 and they differed from macro-Ra values in ranking order. Percentage Gloss values ranged from 30.6 to 70.1%. The results revealed that micro-roughness showed a high correlation with gloss values (r = 0.93), whilst macro-roughness did not (r = 0.62). Moreover, the AFM method showed higher capability to distinguish surface roughness compared with the 2-D profilometry and to reveal more detailed definition of surface texture than the examination under SEM.

Composite Resins↗
PubMed2001

Atomic force microscopy of macromolecular interactions.

The introduction of functional imaging tools and techniques that operate at molecular-length scales has provided investigators with unique approaches to characterizing biomolecular structure and function relationships. Recent advances in the field of scanning probe techniques and, in particular, atomic force microscopy have yielded tantalizing insights into the dynamics of protein self-assembly and the mechanics of protein unfolding.

Crystallography, X-Ray↗
PubMed1997

Atomic force microscopy of collagen molecules. Surface morphology of segment-long-spacing (SLS) crystallites of collagen.

Three-dimensional structures of laterally aggregated bundles of collagen molecules, segment-long-spacing (SLS) crystallites, were imaged by atomic force microscopy (AFM) in atmosphere. The overall image of the type I collagen SLS in a height-amplified mode was semi-cylindrical, approximately 300 nm in length, with two bands of elevation near both N- and C- ends of the molecule. Its 'cross-sectional' profile (across the molecular axis) was a smooth arch. The 'axial' profile (along the molecular axis) had two prominent peaks approximately 250 nm apart, corresponding to the two bands of elevation. There were several minor peaks between these two prominent peaks. The elevation near both ends may be due to the presence of covalently bound sugars near both N- and C- ends of the helical part of type I collagen alpha chains. The AFM images of SLS presented here indicate that the type I collagen molecule is not uniform in diameter and has two bulged parts within its triple helix.

Animals↗
PubMed2004

Atomic force microscopy study of gutta-percha cone topography.

The aim of this study was to investigate the topography of the apical portion of 4 different commercially available standardized gutta-percha (GP) cones with atomic force microscopy (AFM). GP cones (Analytic, Dentsply, Tanari, and Endo Points) were cut 3 mm from the tip, attached in a glass base, and positioned in the AFM. Analyses were performed on 3 different regions of the GP sample and on 3 separate points of each region. Root mean square (RMS) parameters for topographic amplitudes and lateral force variations were calculated. The differences among GP brands were tested by ANOVA with a Fisher protected LSD test of multiple comparisons. According to our results, only Endo Points showed similar behavior between 2 parameters assessed (P <.05). All brands showed normally distributed deep depression in the apical portions, which are within the same scale as proteins and bacterial products that the cones should be blocking. AFM offers a powerful new tool to directly evaluate the characterization of GP cone surfaces.

Analysis of Variance↗
PubMed2005

Assessing transferrin modification of liposomes by atomic force microscopy and transmission electron microscopy.

Site-specific delivery of drugs and therapeutics can significantly reduce drug toxicity and increase the therapeutic effect. Transferrin (Tf) is one suitable ligand to be conjugated to drug delivery systems to achieve site-specific targeting, due to its specific binding to transferrin receptors (TfR), expressed on several cell types of therapeutic interest. TfRs have been reported to be highly expressed on the surfaces of tumour cells and the well-characterised and efficient mechanism of internalisation of Tf has been exploited for the delivery of anticancer drugs, proteins, and therapeutic genes into primarily proliferating malignant cells. Liposomes are effective vehicles for drugs, genes and vaccines and can be easily modified with proteins, antibodies, and other appropriate ligands, resulting in attractive formulations for targeted drug delivery. In this study, we used atomic force microscopy (AFM) and transmission electron microscopy (TEM) to confirm the conjugation of Tf to liposomes by three different coupling methods. In addition, the conventional assays for quantification of protein amount (BCA) and phospholipid content (according to Steward) were performed. AFM and TEM were able to display Tf-molecules on the liposomal surfaces and can be routinely used to obtain additional visual information on the protein-drug carrier conjugation in a fast and reliable manner.

Drug Delivery Systems↗
PubMed2003

Sample preparation and imaging of erythrocyte cytoskeleton with the atomic force microscopy.

A novel method for the covalent attachment of erythrocytes to glass microscope coverslips that can be used to image intact cells and the cytoplasmic side of the cell membrane with either solid or liquid mode atomic force microscopy (AFM) is described. The strong binding of cells to the glass surface is achieved by the interaction of cell membrane carbohydrates to lectin, which is bound to N-5-azido-2-nitrobenzoyloxysuccinimide (ANBNOS)-coated coverslips (1). The effectiveness of this method is compared with the other commonly used methods of immobilizing intact erythrocytes on glass coverslips for AFM observations. Experimental conditions of AFM imaging of biologic tissue are discussed, and typical topographies of the extracellular and the cytoplasmic surfaces of the plasma membrane in the dry state and in the liquid state are presented. Comparison of the spectrin network of cell age-separated erythrocytes has demonstrated significant loss in the network order in older erythrocytes. The changes are quantitatively described using the pixel height histogram and window size grain analysis.

Cell Adhesion↗