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Sample records for afm force spectroscopy

  1. Novel Polymer Linkers for Single Molecule AFM Force Spectroscopy

    PubMed Central

    Tong, Zenghan; Mikheikin, Andrey; Krasnoslobodtsev, Alexey; Lv, Zhengjian; Lyubchenko, Yuri L.

    2013-01-01

    Flexible polymer linkers play an important role in various imaging and probing techniques that require surface immobilization, including atomic force microscopy (AFM). In AFM force spectroscopy, polymer linkers are necessary for the covalent attachment of molecules of interest to the AFM tip and the surface. The polymer linkers tether the molecules and provide their proper orientation in probing experiments. Additionally, the linkers separate specific interactions from nonspecific short-range adhesion and serve as a reference point for the quantitative analysis of single molecule probing events. In this report, we present our results on the synthesis and testing of a novel polymer linker and the identification of a number of potential applications for its use in AFM force spectroscopy experiments. The synthesis of the linker is based on the well-developed phosphoramidate (PA) chemistry that allows the routine synthesis of linkers with predetermined lengths and PA composition. These linkers are homogeneous in length and can be terminated with various functional groups. PA linkers with different functional groups were synthesized and tested in experimental systems utilizing different immobilization chemistries. We probed interactions between complementary DNA oligonucleotides; DNA and protein complexes formed by the site-specific binding protein SfiI; and interactions between amyloid peptide (Aβ42). The results of the AFM force spectroscopy experiments validated the feasibility of the proposed approach for the linker design and synthesis. Furthermore, the properties of the tether (length, functional groups) can be adjusted to meet the specific requirements for different force spectroscopy experiments and system characteristics, suggesting that it could be used for a large number of various applications. PMID:23624104

  2. Single-Molecule Studies of Integrins by AFM-Based Force Spectroscopy on Living Cells

    NASA Astrophysics Data System (ADS)

    Eibl, Robert H.

    The characterization of cell adhesion between two living cells at the single-molecule level, i.e., between one adhesion receptor and its counter-receptor, appears to be an experimental challenge. Atomic force microscopy (AFM) can be used in its force spectroscopy mode to determine unbinding forces of a single pair of adhesion receptors, even with a living cell as a probe. This chapter provides an overview of AFM force measurements of the integrin family of cell adhesion receptors and their ligands. A focus is given to major integrins expressed on leukocytes, such as lymphocyte function-associated antigen 1 (LFA-1) and very late antigen 4 (VLA-4). These receptors are crucial for leukocyte trafficking in health and disease. LFA-1 and VLA-1 can be activated within the bloodstream from a low-affinity to a high-affinity receptor by chemokines in order to adhere strongly to the vessel wall before the receptor-bearing leukocytes extravasate. The experimental considerations needed to provide near-physiological conditions for a living cell and to be able to measure adequate forces at the single-molecule level are discussed in detail. AFM technology has been developed into a modern and extremely sensitive tool in biomedical research. It appears now that AFM force spectroscopy could enter, within a few years, medical applications in diagnosis and therapy of cancer and autoimmune diseases.

  3. Study on the AFM Force Spectroscopy method for elastic modulus measurement of living cells

    NASA Astrophysics Data System (ADS)

    Demichelis, A.; Pavarelli, S.; Mortati, L.; Sassi, G.; Sassi, M.

    2013-09-01

    The cell elasticity gives information about its pathological state and metastatic potential. The aim of this paper is to study the AFM Force Spectroscopy technique with the future goal of realizing a reference method for accurate elastic modulus measurement in the elasticity range of living cells. This biological range has not been yet explored with a metrological approach. Practical hints are given for the realization of a Sylgard elasticity scale. Systematic effects given by the sample curing thickness and nanoindenter geometry have been found with regards of the measured elastic modulus. AFM measurement reproducibility better than 20% is obtained in the entire investigated elastic modulus scale of 101 - 104 kPa.

  4. Nanopuller-open data acquisition platform for AFM force spectroscopy experiments.

    PubMed

    Pawlak, Konrad; Strzelecki, Janusz

    2016-05-01

    Atomic Force Microscope (AFM) is a widely used tool in force spectroscopy studies. Presently, this instrument is accessible from numerous vendors, albeit commercial solutions are expensive and almost always hardware and software closed. Approaches for open setups were published, as with modern low cost and readily available piezoelectric actuators, data acquisition interfaces and optoelectronic components building such force spectroscopy AFM is relatively easy. However, suitable software to control such laboratory made instrument was not released. Developing it in the lab requires significant time and effort. Our Nanopuller software described in this paper is intended to eliminate this obstacle. With only minimum adjustments this program can be used to control and acquire data with any suitable National Instruments universal digital/analog interface and piezoelectric actuator analog controller, giving significant freedom and flexibility in designing force spectroscopy experiment. Since the full code, written in a graphical LabVIEW environment is available, our Nanopuller can be easily customized. In this paper we describe the program and test its performance in controlling different setups. Successful and accurate force curve acquisition for standard samples (single molecules of I27O reference titin polyprotein and DNA as well as red blood cells) is shown.

  5. Structural impact of cations on lipid bilayer models: nanomechanical properties by AFM-force spectroscopy.

    PubMed

    Redondo-Morata, Lorena; Giannotti, Marina I; Sanz, Fausto

    2014-02-01

    Atomic Force Microscopy (AFM) has become an invaluable tool for studying the micro- and nanoworlds. As a stand-alone, high-resolution imaging technique and force transducer, it defies most other surface instrumentation in ease of use, sensitivity and versatility. The main strength of AFM relies on the possibility to operate in an aqueous environment on a wide variety of biological samples, from single molecules - DNA or proteins - to macromolecular assemblies like biological membranes. Understanding the effect of mechanical stress on membranes is of primary importance in biophysics, since cells are known to perform their function under a complex combination of forces. In the later years, AFM-based Force-Spectroscopy (AFM-FS) has provided a new vista on membrane mechanics in a confined area within the nanometer realm, where most of the specific molecular interactions take place. Lipid membranes are electrostatically charged entities that physiologically coexist with electrolyte solutions. Thus, specific interactions with ions are a matter of considerable interest. The distribution of ions in the solution and their interaction with the membranes are factors that substantially modify the structure and dynamics of the cell membranes. Furthermore, signaling processes are modified by the membrane capability of retaining ions. Supported Lipid Bilayers (SLBs) are a versatile tool to investigate phospholipid membranes mimicking biological surfaces. In the present contribution, we review selected experiments on the mechanical stability of SLBs as models of lipid membranes by means of AFM-FS, with special focus on the effect of cations and ionic strength in the overall nanomechanical stability. PMID:24341385

  6. Plant protein interactions studied using AFM force spectroscopy: nanomechanical and adhesion properties.

    PubMed

    Fahs, Ahmad; Louarn, Guy

    2013-07-21

    The present work was focused on the nanomechanical and adhesion properties of the napin (2S albumin) and cruciferin (12S globulin) rapeseed (Brassica napus L.) proteins, respectively, a low and high molecular weight seed protein. Using chemically modified AFM tips, force spectroscopy experiments demonstrated notable differences in the tip-protein interaction strength with regard to the nature of the protein and pH of the aqueous environment. The results clearly underline the role of residence time and electrostatic interactions in the protein-protein adhesion force. Although the nanomechanical experiments concerned more than a single molecule, unfolding length and force characteristics of the rapeseed proteins have been statistically found to be sensitive to the structural properties of the protein. This study provides insight into the characterization of rapeseed proteins and then a better knowledge of their interaction and assembling at the nanoscale range.

  7. Plant protein interactions studied using AFM force spectroscopy: nanomechanical and adhesion properties.

    PubMed

    Fahs, Ahmad; Louarn, Guy

    2013-07-21

    The present work was focused on the nanomechanical and adhesion properties of the napin (2S albumin) and cruciferin (12S globulin) rapeseed (Brassica napus L.) proteins, respectively, a low and high molecular weight seed protein. Using chemically modified AFM tips, force spectroscopy experiments demonstrated notable differences in the tip-protein interaction strength with regard to the nature of the protein and pH of the aqueous environment. The results clearly underline the role of residence time and electrostatic interactions in the protein-protein adhesion force. Although the nanomechanical experiments concerned more than a single molecule, unfolding length and force characteristics of the rapeseed proteins have been statistically found to be sensitive to the structural properties of the protein. This study provides insight into the characterization of rapeseed proteins and then a better knowledge of their interaction and assembling at the nanoscale range. PMID:23732983

  8. Functionalized AFM probes for force spectroscopy: eigenmode shapes and stiffness calibration through thermal noise measurements.

    PubMed

    Laurent, Justine; Steinberger, Audrey; Bellon, Ludovic

    2013-06-01

    The functionalization of an atomic force microscope (AFM) cantilever with a colloidal bead is a widely used technique when the geometry between the probe and the sample must be controlled, particularly in force spectroscopy. But some questions remain: how does a bead glued at the end of a cantilever influence its mechanical response? And more importantly for quantitative measurements, can we still determine the stiffness of the AFM probe with traditional techniques?In this paper, the influence of the colloidal mass loading on the eigenmode shape and resonant frequency is investigated by measuring the thermal noise on rectangular AFM microcantilevers with and without beads attached at their extremities. The experiments are performed with a home-made ultra-sensitive AFM, based on differential interferometry. The focused beam from the interferometer probes the cantilever at different positions and the spatial shapes of the modes are determined up to the fifth resonance, without external excitation. The results clearly demonstrate that the first eigenmode is almost unchanged by mass loading. However the oscillation behavior of higher resonances presents a marked difference: with a particle glued at its extremity, the nodes of the modes are displaced towards the free end of the cantilever. These results are compared to an analytical model taking into account the mass and inertial moment of the load in an Euler-Bernoulli framework, where the normalization of the eigenmodes is explicitly worked out in order to allow a quantitative prediction of the thermal noise amplitude of each mode. A good agreement between the experimental results and the analytical model is demonstrated, allowing a clean calibration of the probe stiffness.

  9. Unspecific membrane protein-lipid recognition: combination of AFM imaging, force spectroscopy, DSC and FRET measurements.

    PubMed

    Borrell, Jordi H; Montero, M Teresa; Morros, Antoni; Domènech, Òscar

    2015-11-01

    In this work, we will describe in quantitative terms the unspecific recognition between lactose permease (LacY) of Escherichia coli, a polytopic model membrane protein, and one of the main components of the inner membrane of this bacterium. Supported lipid bilayers of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG) (3:1, mol/mol) in the presence of Ca(2+) display lateral phase segregation that can be distinguished by atomic force microscopy (AFM) as well as force spectroscopy. LacY shows preference for fluid (Lα) phases when it is reconstituted in POPE : POPG (3:1, mol/mol) proteoliposomes at a lipid-to-protein ratio of 40. When the lipid-to-protein ratio is decreased down to 0.5, two domains can be distinguished by AFM. While the upper domain is formed by self-segregated units of LacY, the lower domain is constituted only by phospholipids in gel (Lβ) phase. On the one hand, classical differential scanning calorimetry (DSC) measurements evidenced the segregation of a population of phospholipids and point to the existence of a boundary region at the lipid-protein interface. On the other hand, Förster Resonance Energy Transfer (FRET) measurements in solution evidenced that POPE is selectively recognized by LacY. A binary pseudophase diagram of POPE : POPG built from AFM observations enables to calculate the composition of the fluid phase where LacY is inserted. These results are consistent with a model where POPE constitutes the main component of the lipid-LacY interface segregated from the fluid bulk phase where POPG predominates.

  10. The physics of pulling polyproteins: a review of single molecule force spectroscopy using the AFM to study protein unfolding

    NASA Astrophysics Data System (ADS)

    Hughes, Megan L.; Dougan, Lorna

    2016-07-01

    One of the most exciting developments in the field of biological physics in recent years is the ability to manipulate single molecules and probe their properties and function. Since its emergence over two decades ago, single molecule force spectroscopy has become a powerful tool to explore the response of biological molecules, including proteins, DNA, RNA and their complexes, to the application of an applied force. The force versus extension response of molecules can provide valuable insight into its mechanical stability, as well as details of the underlying energy landscape. In this review we will introduce the technique of single molecule force spectroscopy using the atomic force microscope (AFM), with particular focus on its application to study proteins. We will review the models which have been developed and employed to extract information from single molecule force spectroscopy experiments. Finally, we will end with a discussion of future directions in this field.

  11. The physics of pulling polyproteins: a review of single molecule force spectroscopy using the AFM to study protein unfolding.

    PubMed

    Hughes, Megan L; Dougan, Lorna

    2016-07-01

    One of the most exciting developments in the field of biological physics in recent years is the ability to manipulate single molecules and probe their properties and function. Since its emergence over two decades ago, single molecule force spectroscopy has become a powerful tool to explore the response of biological molecules, including proteins, DNA, RNA and their complexes, to the application of an applied force. The force versus extension response of molecules can provide valuable insight into its mechanical stability, as well as details of the underlying energy landscape. In this review we will introduce the technique of single molecule force spectroscopy using the atomic force microscope (AFM), with particular focus on its application to study proteins. We will review the models which have been developed and employed to extract information from single molecule force spectroscopy experiments. Finally, we will end with a discussion of future directions in this field.

  12. Force spectroscopy 101: how to design, perform, and analyze an AFM-based single molecule force spectroscopy experiment.

    PubMed

    Noy, Aleksandr

    2011-10-01

    Single molecule force spectroscopy presents a deceptively simple approach to probing interaction between molecules and molecular assemblies on the nanoscale by measuring forces that it takes to pull the molecules apart. Yet, a more detailed analysis reveals a wealth of different behaviors and interesting physics. This article aims to explore basic physical concepts behind these experiments from a strictly practical point of using these data to extract meaningful information about the interactions. It also focuses on different loading regimes in these experiments, different kinetics that they cause, and different data interpretation that is required for measurements in those regimes. PMID:21862386

  13. Measuring protein isoelectric points by AFM-based force spectroscopy using trace amounts of sample.

    PubMed

    Guo, Shifeng; Zhu, Xiaoying; Jańczewski, Dominik; Lee, Serina Siew Chen; He, Tao; Teo, Serena Lay Ming; Vancso, G Julius

    2016-09-01

    Protein charge at various pH and isoelectric point (pI) values is important in understanding protein function. However, often only trace amounts of unknown proteins are available and pI measurements cannot be obtained using conventional methods. Here, we show a method based on the atomic force microscope (AFM) to determine pI using minute quantities of proteins. The protein of interest is immobilized on AFM colloidal probes and the adhesion force of the protein is measured against a positively and a negatively charged substrate made by layer-by-layer deposition of polyelectrolytes. From the AFM force-distance curves, pI values with an estimated accuracy of ±0.25 were obtained for bovine serum albumin, myoglobin, fibrinogen and ribonuclease A over a range of 4.7-9.8. Using this method, we show that the pI of the 'footprint' of the temporary adhesive proteins secreted by the barnacle cyprid larvae of Amphibalanus amphitrite is in the range 9.6-9.7. PMID:27454881

  14. Measuring protein isoelectric points by AFM-based force spectroscopy using trace amounts of sample

    NASA Astrophysics Data System (ADS)

    Guo, Shifeng; Zhu, Xiaoying; Jańczewski, Dominik; Lee, Serina Siew Chen; He, Tao; Teo, Serena Lay Ming; Vancso, G. Julius

    2016-09-01

    Protein charge at various pH and isoelectric point (pI) values is important in understanding protein function. However, often only trace amounts of unknown proteins are available and pI measurements cannot be obtained using conventional methods. Here, we show a method based on the atomic force microscope (AFM) to determine pI using minute quantities of proteins. The protein of interest is immobilized on AFM colloidal probes and the adhesion force of the protein is measured against a positively and a negatively charged substrate made by layer-by-layer deposition of polyelectrolytes. From the AFM force-distance curves, pI values with an estimated accuracy of ±0.25 were obtained for bovine serum albumin, myoglobin, fibrinogen and ribonuclease A over a range of 4.7-9.8. Using this method, we show that the pI of the ‘footprint’ of the temporary adhesive proteins secreted by the barnacle cyprid larvae of Amphibalanus amphitrite is in the range 9.6-9.7.

  15. Bacteria attachment to surfaces--AFM force spectroscopy and physicochemical analyses.

    PubMed

    Harimawan, Ardiyan; Rajasekar, Aruliah; Ting, Yen-Peng

    2011-12-01

    Understanding bacterial adhesion to surfaces requires knowledge of the forces that govern bacterial-surface interactions. Biofilm formation on stainless steel 316 (SS316) by three bacterial species was investigated by examining surface force interaction between the cells and metal surface using atomic force microscopy (AFM). Bacterial-metal adhesion force was quantified at different surface delay time from 0 to 60s using AFM tip coated with three different bacterial species: Gram-negative Massilia timonae and Pseudomonas aeruginosa, and Gram-positive Bacillus subtilis. The results revealed that bacterial adhesion forces on SS316 surface by Gram-negative bacteria is higher (8.53±1.40 nN and 7.88±0.94 nN) when compared to Gram-positive bacteria (1.44±0.21 nN). Physicochemical analysis on bacterial surface properties also revealed that M. timonae and P. aeruginosa showed higher hydrophobicity and surface charges than B. subtilis along with the capability of producing extracellular polymeric substances (EPS). The higher hydrophobicity, surface charges, and greater propensity to form EPS by M. timonae and P. aeruginosa led to high adhesive force on the metal surface. PMID:21889162

  16. Measuring protein isoelectric points by AFM-based force spectroscopy using trace amounts of sample

    NASA Astrophysics Data System (ADS)

    Guo, Shifeng; Zhu, Xiaoying; Jańczewski, Dominik; Lee, Serina Siew Chen; He, Tao; Teo, Serena Lay Ming; Vancso, G. Julius

    2016-09-01

    Protein charge at various pH and isoelectric point (pI) values is important in understanding protein function. However, often only trace amounts of unknown proteins are available and pI measurements cannot be obtained using conventional methods. Here, we show a method based on the atomic force microscope (AFM) to determine pI using minute quantities of proteins. The protein of interest is immobilized on AFM colloidal probes and the adhesion force of the protein is measured against a positively and a negatively charged substrate made by layer-by-layer deposition of polyelectrolytes. From the AFM force–distance curves, pI values with an estimated accuracy of ±0.25 were obtained for bovine serum albumin, myoglobin, fibrinogen and ribonuclease A over a range of 4.7–9.8. Using this method, we show that the pI of the ‘footprint’ of the temporary adhesive proteins secreted by the barnacle cyprid larvae of Amphibalanus amphitrite is in the range 9.6–9.7.

  17. Tip-surface interactions at redox responsive poly(ferrocenylsilane) (PFS) interface by AFM-based force spectroscopy

    NASA Astrophysics Data System (ADS)

    Chung, Hong Jing; Song, Jing; Vancso, G. Julius

    2009-05-01

    Poly(ferrocenylsilanes) (PFS) belong to the class of redox responsive organometallic polymers. Atomic force microscopy (AFM)-based single molecule force spectroscopy (SMFS) was used earlier to study single chain PFS response and redox energy driven single chain PFS molecular motors. Here we present further AFM investigations of force interactions between tip and a grafted PFS surface under potential control in electrochemical redox cycles. Typical tip-Au interaction is considered as reference in the force measurements. First the electrostatic component in the diffused double layer (DL) in NaClO 4 electrolyte environment was considered for a "grafted to" PFS, which dominated the interplay between the tip and sample surface. The DL forces can also hinder the physisorption of PFS chain onto the tip when the voltage was applied at -0.1 V. On the other hand, if the tip contacted the PFS surface prior to the electrochemical process, physisorption of PFS chains governed the overall interaction regardless of subsequently applied surface potential. In addition, prolonged contact time, tc, may also contribute to the stability of tip-PFS bridging and detection of electrostatic forces between the tip-PFS interface. The results showed that tip-substrate interaction forces without PFS grafts have negligibly small force contributions under similar, electrochemically controlled, conditions used in single PFS chain based molecular motors.

  18. Experimental evidence of ultrathin polymer film stratification by AFM force spectroscopy.

    PubMed

    Delorme, Nicolas; Chebil, Mohamed Souheib; Vignaud, Guillaume; Le Houerou, Vincent; Bardeau, Jean-François; Busselez, Rémi; Gibaud, Alain; Grohens, Yves

    2015-06-01

    By performing Atomic Force Microscopy measurements of pull-off force as a function of the temperature, we were able to probe the dynamic of supported thin polystyrene (PS) films. Thermal transitions induce modifications in the surface energy, roughness and surface modulus that are clearly detected by AFM and related to PS chain relaxation mechanisms. We demonstrated the existence of three transition temperatures that can be associated to the relaxation of polymer chains located at different depth regions within the polymer film. Independently of the film thickness, we have confirmed the presence of a region of high mobility for the polymer chains at the free interface. The thickness of this region is estimated to be above 7nm. The detection of a transition only present for film thicker than the gyration radius Rg is linked to the dynamics of polymer chains in a bulk conformation (i.e. not in contact with the free interface). We claim here that our results demonstrate, in agreement with other techniques, the stratification of thin polymer film depth profile in terms of relaxation behavior. PMID:26087914

  19. Mechanical properties of complex biological systems using AFM-based force spectroscopy

    NASA Astrophysics Data System (ADS)

    Graham, John Stephen

    An atomic force microscope (AFM) was designed and built to study the mechanical properties of small collagen fibrils and the plasma membrane of living cells. Collagen is a major component of bone, skin and connective tissues, and is abundant in the extracellular matrix (ECM). Because of its abundance, an understanding of how disease affects collagen mechanics is crucial in disease prevention efforts. Two levels of type I collagen structure were investigated, subfibrils (on the order of 1 mum in length) and longer fibrils. Comparisons were made between measurements of wild-type (wt) collagen and collagen from the mouse model of osteogenesis imperfecta (OI). Significant differences between OI and wt collagen were observed, primarily that intermolecular bonds in OI collagen fibrils are weaker than in wt, or not ruptured, as in the case of OI subfibrils. As cells interact with collagen in the ECM, the mechanical properties of the plasma membrane are also of great interest. Membrane tethers were extracted from living cells under varied conditions in order to assess the contributions of membrane-associated macromolecules such as the actin cytoskeleton and the glycocalyx, and intracellular signaling. Tether extraction force was found to be sensitive to all of these altered conditions, suggesting that tether extraction may be used to monitor various cellular processes.

  20. Impact of galactosylceramides on the nanomechanical properties of lipid bilayer models: an AFM-force spectroscopy study.

    PubMed

    Gumí-Audenis, Berta; Sanz, Fausto; Giannotti, Marina I

    2015-07-21

    Galactosylceramides (GalCer) are glycosphingolipids bound to a monosaccharide group, responsible for inducing extensive hydrogen bonds that yield their alignment and accumulation in the outer leaflet of the biological membrane together with cholesterol (Chol) in rafts. In this work, the influence of GalCer on the nanomechanical properties of supported lipid bilayers (SLBs) based on DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) and DLPC (1,2-didodecanoyl-sn-glycero-3-phosphocoline) as model systems was assessed. Phosphatidylcholine (PC):GalCer SLBs were characterized by means of differential scanning calorimetry (DSC) and atomic force microscopy (AFM), in both imaging and force spectroscopy (AFM-FS) modes. Comparing both PC systems, we determined that the behaviour of SLB mixtures is governed by the PC phase-like state at the working temperature. While a phase segregated system is observed for DLPC:GalCer SLBs, GalCer are found to be dissolved in DPPC SLBs for GalCer contents up to 20 mol%. In both systems, the incorporation of GalCer intensifies the nanomechanical properties of SLBs. Interestingly, segregated domains of exceptionally high mechanical stability are formed in DLPC:GalCer SLBs. Finally, the role of 20 mol% Chol in GalCer organization and function in the membranes was assessed. Both PC model systems displayed phase segregation and remarkable nanomechanical stability when GalCer and Chol coexist in SLBs. PMID:26058499

  1. Complete noise analysis of a simple force spectroscopy AFM setup and its applications to study nanomechanics of mammalian Notch 1 protein

    NASA Astrophysics Data System (ADS)

    Dey, Ashim; Szoszkiewicz, Robert

    2012-05-01

    We describe a complete noise analysis and application of a custom made AFM force spectroscopy setup on pulling a recombinant protein with an NRR domain of mouse Notch 1. Our table top AFM setup is affordable, has an open architecture, and is easily transferable to other laboratories. Its calculated noise characteristics are dominated by the Brownian noise with 2% non-Brownian components integrated over the first thermally induced resonance of a typical cantilever. For a typical SiN cantilever with a force constant of ˜15 pN nm-1 and in water the force sensitivity and resolution are less than 10 pN, and the corresponding deflection sensitivities are less than 100 pm Hz-1/2. Also, we obtain a sub-ms time resolution in detecting the protein length change, and only few ms cantilever response times as measured in the force clamp mode on a well-known protein standard. Using this setup we investigate force-induced conformational transitions in the NRR region of a mouse Notch 1. Notch is an important protein related to leukemia and breast cancers in humans. We demonstrate that it is feasible to develop AFM-based studies of the force-induced conformational transitions in Notch. Our results match recent steered molecular dynamics simulations of the NRR unfolding and constitute a first step towards a detailed study of Notch activation with AFM.

  2. Measurement of membrane binding between recoverin, a calcium-myristoyl switch protein, and lipid bilayers by AFM-based force spectroscopy.

    PubMed Central

    Desmeules, Philippe; Grandbois, Michel; Bondarenko, Vladimir A; Yamazaki, Akio; Salesse, Christian

    2002-01-01

    Myristoyl switch is a feature of several peripheral membrane proteins involved in signal transduction pathways. This unique molecular property is best illustrated by the "Ca(2+)-myristoyl switch" of recoverin, which is a Ca(2+)-binding protein present in retinal rod cells of vertebrates. In this transduction pathway, the Ca(2+)-myristoyl switch acts as a calcium sensor involved in cell recovery from photoactivation. Ca(2+) binding by recoverin induces the extrusion of its myristoyl group to the solvent, which leads to its translocation from cytosol to rod disk membranes. Force spectroscopy, based on atomic force microscope (AFM) technology, was used to determine the extent of membrane binding of recoverin in the absence and presence of calcium, and to quantify this force of binding. An adhesion force of 48 +/- 5 pN was measured between recoverin and supported phospholipid bilayers in the presence of Ca(2+). However, no binding was observed in the absence of Ca(2+). Experiments with nonmyristoylated recoverin confirmed these observations. Our results are consistent with previously measured extraction forces of lipids from membranes. PMID:12023256

  3. AFM force spectroscopy reveals how subtle structural differences affect the interaction strength between Candida albicans and DC-SIGN.

    PubMed

    te Riet, Joost; Reinieren-Beeren, Inge; Figdor, Carl G; Cambi, Alessandra

    2015-11-01

    The fungus Candida albicans is the most common cause of mycotic infections in immunocompromised hosts. Little is known about the initial interactions between Candida and immune cell receptors, such as the C-type lectin dendritic cell-specific intracellular cell adhesion molecule-3 (ICAM-3)-grabbing non-integrin (DC-SIGN), because a detailed characterization at the structural level is lacking. DC-SIGN recognizes specific Candida-associated molecular patterns, that is, mannan structures present in the cell wall of Candida. The molecular recognition mechanism is however poorly understood. We postulated that small differences in mannan-branching may result in considerable differences in the binding affinity. Here, we exploit atomic force microscope-based dynamic force spectroscopy with single Candida cells to gain better insight in the carbohydrate recognition capacity of DC-SIGN. We demonstrate that slight differences in the N-mannan structure of Candida, that is, the absence or presence of a phosphomannan side chain, results in differences in the recognition by DC-SIGN as follows: (i) it contributes to the compliance of the outer cell wall of Candida, and (ii) its presence results in a higher binding energy of 1.6 kB T. The single-bond affinity of tetrameric DC-SIGN for wild-type C. albicans is ~10.7 kB T and a dissociation constant kD of 23 μM, which is relatively strong compared with other carbohydrate-protein interactions described in the literature. In conclusion, this study shows that DC-SIGN specifically recognizes mannan patterns on C. albicans with high affinity. Knowledge on the binding pocket of DC-SIGN and its pathogenic ligands will lead to a better understanding of how fungal-associated carbohydrate structures are recognized by receptors of the immune system and can ultimately contribute to the development of new anti-fungal drugs.

  4. Hydrodynamic effects in fast AFM single-molecule force measurements.

    PubMed

    Janovjak, Harald; Struckmeier, Jens; Müller, Daniel J

    2005-02-01

    Atomic force microscopy (AFM) allows the critical forces that unfold single proteins and rupture individual receptor-ligand bonds to be measured. To derive the shape of the energy landscape, the dynamic strength of the system is probed at different force loading rates. This is usually achieved by varying the pulling speed between a few nm/s and a few microm/s, although for a more complete investigation of the kinetic properties higher speeds are desirable. Above 10 microm/s, the hydrodynamic drag force acting on the AFM cantilever reaches the same order of magnitude as the molecular forces. This has limited the maximum pulling speed in AFM single-molecule force spectroscopy experiments. Here, we present an approach for considering these hydrodynamic effects, thereby allowing a correct evaluation of AFM force measurements recorded over an extended range of pulling speeds (and thus loading rates). To support and illustrate our theoretical considerations, we experimentally evaluated the mechanical unfolding of a multi-domain protein recorded at 30 microm/s pulling speed. PMID:15257425

  5. Near-Field Spectroscopy with Nanoparticles Deposited by AFM

    NASA Technical Reports Server (NTRS)

    Anderson, Mark S.

    2008-01-01

    An alternative approach to apertureless near-field optical spectroscopy involving an atomic-force microscope (AFM) entails less complexity of equipment than does a prior approach. The alternative approach has been demonstrated to be applicable to apertureless near-field optical spectroscopy of the type using an AFM and surface enhanced Raman scattering (SERS), and is expected to be equally applicable in cases in which infrared or fluorescence spectroscopy is used. Apertureless near-field optical spectroscopy is a means of performing spatially resolved analyses of chemical compositions of surface regions of nanostructured materials. In apertureless near-field spectroscopy, it is common practice to utilize nanostructured probe tips or nanoparticles (usually of gold) having shapes and dimensions chosen to exploit plasmon resonances so as to increase spectroscopic-signal strengths. To implement the particular prior approach to which the present approach is an alternative, it is necessary to integrate a Raman spectrometer with an AFM and to utilize a special SERS-active probe tip. The resulting instrumentation system is complex, and the tasks of designing and constructing the system and using the system to acquire spectro-chemical information from nanometer-scale regions on a surface are correspondingly demanding.

  6. ezAFM: A low cost Atomic Force Microscope(AFM)

    NASA Astrophysics Data System (ADS)

    Celik, Umit; Celik, Kubra; Aslan, Husnu; Kehribar, Ihsan; Dede, Munir; Ozgur Ozer, H.; Oral, Ahmet

    2012-02-01

    A low cost AFM, ezAFM is developed for educational purposes as well as research. Optical beam deflection method is used to measure the deflection of cantilever. ezAFM scanner is built using voice coil motors (VCM) with ˜50x50x6 μm scan area. The microscope uses alignment free cantilevers, which minimizes setup times. FPGA based AFM feedback Control electronics is developed. FPGA technology allows us to drive all peripherals in parallel. ezAFM Controller is connected to PC by USB 2.0 interface as well as Wi-Fi. We have achieved <5nm lateral and ˜0.01nm vertical resolution. ezAFM can image single atomic steps in HOPG and mica. An optical microscope with <3 μm resolution is also integrated into the system. ezAFM supports different AFM operation modes such as dynamic mode, contact mode, lateral force microscopy. Advanced modes like magnetic force microscopy and electric force microscopy will be implemented later on. The new ezAFM system provides, short learning times for student labs, quick setup and easy to transport for portable applications with the best price/performance ratio. The cost of the system starts from 15,000, with system performance comparable with the traditional AFM systems.

  7. In situ hydrodynamic lateral force calibration of AFM colloidal probes.

    PubMed

    Ryu, Sangjin; Franck, Christian

    2011-11-01

    Lateral force microscopy (LFM) is an application of atomic force microscopy (AFM) to sense lateral forces applied to the AFM probe tip. Recent advances in tissue engineering and functional biomaterials have shown a need for the surface characterization of their material and biochemical properties under the application of lateral forces. LFM equipped with colloidal probes of well-defined tip geometries has been a natural fit to address these needs but has remained limited to provide primarily qualitative results. For quantitative measurements, LFM requires the successful determination of the lateral force or torque conversion factor of the probe. Usually, force calibration results obtained in air are used for force measurements in liquids, but refractive index differences between air and liquids induce changes in the conversion factor. Furthermore, in the case of biochemically functionalized tips, damage can occur during calibration because tip-surface contact is inevitable in most calibration methods. Therefore, a nondestructive in situ lateral force calibration is desirable for LFM applications in liquids. Here we present an in situ hydrodynamic lateral force calibration method for AFM colloidal probes. In this method, the laterally scanned substrate surface generated a creeping Couette flow, which deformed the probe under torsion. The spherical geometry of the tip enabled the calculation of tip drag forces, and the lateral torque conversion factor was calibrated from the lateral voltage change and estimated torque. Comparisons with lateral force calibrations performed in air show that the hydrodynamic lateral force calibration method enables quantitative lateral force measurements in liquid using colloidal probes.

  8. Introduction to Atomic Force Microscopy (AFM) in Biology.

    PubMed

    Kreplak, Laurent

    2016-01-01

    The atomic force microscope (AFM) has the unique capability of imaging biological samples with molecular resolution in buffer solution over a wide range of time scales from milliseconds to hours. In addition to providing topographical images of surfaces with nanometer- to angstrom-scale resolution, forces between single molecules and mechanical properties of biological samples can be investigated from the nano-scale to the micro-scale. Importantly, the measurements are made in buffer solutions, allowing biological samples to "stay alive" within a physiological-like environment while temporal changes in structure are measured-e.g., before and after addition of chemical reagents. These qualities distinguish AFM from conventional imaging techniques of comparable resolution, e.g., electron microscopy (EM). This unit provides an introduction to AFM on biological systems and describes specific examples of AFM on proteins, cells, and tissues. The physical principles of the technique and methodological aspects of its practical use and applications are also described. © 2016 by John Wiley & Sons, Inc. PMID:27479503

  9. The importance of correcting for variable probe-sample interactions in AFM-IR spectroscopy: AFM-IR of dried bacteria on a polyurethane film.

    PubMed

    Barlow, Daniel E; Biffinger, Justin C; Cockrell-Zugell, Allison L; Lo, Michael; Kjoller, Kevin; Cook, Debra; Lee, Woo Kyung; Pehrsson, Pehr E; Crookes-Goodson, Wendy J; Hung, Chia-Suei; Nadeau, Lloyd J; Russell, John N

    2016-08-01

    AFM-IR is a combined atomic force microscopy-infrared spectroscopy method that shows promise for nanoscale chemical characterization of biological-materials interactions. In an effort to apply this method to quantitatively probe mechanisms of microbiologically induced polyurethane degradation, we have investigated monolayer clusters of ∼200 nm thick Pseudomonas protegens Pf-5 bacteria (Pf) on a 300 nm thick polyether-polyurethane (PU) film. Here, the impact of the different biological and polymer mechanical properties on the thermomechanical AFM-IR detection mechanism was first assessed without the additional complication of polymer degradation. AFM-IR spectra of Pf and PU were compared with FTIR and showed good agreement. Local AFM-IR spectra of Pf on PU (Pf-PU) exhibited bands from both constituents, showing that AFM-IR is sensitive to chemical composition both at and below the surface. One distinct difference in local AFM-IR spectra on Pf-PU was an anomalous ∼4× increase in IR peak intensities for the probe in contact with Pf versus PU. This was attributed to differences in probe-sample interactions. In particular, significantly higher cantilever damping was observed for probe contact with PU, with a ∼10× smaller Q factor. AFM-IR chemical mapping at single wavelengths was also affected. We demonstrate ratioing of mapping data for chemical analysis as a simple method to cancel the extreme effects of the variable probe-sample interactions. PMID:27403761

  10. Study of relaxation and transport processes by means of AFM based dielectric spectroscopy

    SciTech Connect

    Miccio, Luis A.

    2014-05-15

    Since its birth a few years ago, dielectric spectroscopy studies based on atomic force microscopy (AFM) have gained a growing interest. Not only the frequency and temperature ranges have become broader since then but also the kind of processes that can be studied by means of this approach. In this work we analyze the most adequate experimental setup for the study of several dielectric processes with a spatial resolution of a few nanometers by using force mode AFM based dielectric spectroscopy. Proof of concept experiments were performed on PS/PVAc blends and PMMA homopolymer films, for temperatures ranging from 300 to 400 K. Charge transport processes were also studied by this approach. The obtained results were analyzed in terms of cantilever stray contribution, film thickness and relaxation strength. We found that the method sensitivity is strongly coupled with the film thickness and the relaxation strength, and that it is possible to control it by using an adequate experimental setup.

  11. AFM-based force microsensor for a microrobot

    NASA Astrophysics Data System (ADS)

    Fatikow, Sergej; Fahlbusch, Stephan

    2001-10-01

    Microrobots are the result of increasing research activities at the border between microsystem technology and robotics. Today already, robots with dimensions of a few cubic- centimeters can be developed. Like conventional robots, microrobots represent a complex system that usually contains several different types of actuators and sensors. The measurement of gripping forces is the most important sensor application in micromanipulation besides visual servoing to protect the parts from too high surface pressures and thereby damage during the assembly process. Very small forces in the range of 200 (mu) N down to 0.1 (mu) N or even less have to be sensed. Thus, the aim of our current research activities is the development of a high-resolution integrated force microsensor for measuring gripping forces in a microhandling robot. On the one hand, the sensor should be a device for teleoperated manipulation tasks in a flexible microhandling station. On the other hand, typical microhandling operations should to a large extend be automated with the aid of computer-based signal processing of sensor information. The user should be provided with an interface for teleoperated manipulation and an interface for partially automated manipulation of microobjects. In this paper, a concept for the measurement of gripping forces in microrobotics using piezoresistive AFM (atomic force microscope) cantilevers is introduced. Further on, the concept of a microrobot-based SEM station and its applications are presented.

  12. Atomic Force Microscope for Imaging and Spectroscopy

    NASA Technical Reports Server (NTRS)

    Pike, W. T.; Hecht, M. H.; Anderson, M. S.; Akiyama, T.; Gautsch, S.; deRooij, N. F.; Staufer, U.; Niedermann, Ph.; Howald, L.; Mueller, D.

    2000-01-01

    We have developed, built, and tested an atomic force microscope (AFM) for extraterrestrial applications incorporating a micromachined tip array to allow for probe replacement. It is part of a microscopy station originally intended for NASA's 2001 Mars lander to identify the size, distribution, and shape of Martian dust and soil particles. As well as imaging topographically down to nanometer resolution, this instrument can be used to reveal chemical information and perform infrared and Raman spectroscopy at unprecedented resolution.

  13. A review of the application of atomic force microscopy (AFM) in food science and technology.

    PubMed

    Liu, Shaoyang; Wang, Yifen

    2011-01-01

    Atomic force microscopy (AFM) is a powerful nanoscale analysis technique used in food area. This versatile technique can be used to acquire high-resolution sample images and investigate local interactions in air or liquid surroundings. In this chapter, we explain the principles of AFM and review representative applications of AFM in gelatin, casein micelle, carrageenan, gellan gum, starch, and interface. We elucidate new knowledge revealed with AFM as well as ways to use AFM to obtain morphology and rheology information in different food fields.

  14. A detailed guideline for the fabrication of single bacterial probes used for atomic force spectroscopy.

    PubMed

    Thewes, Nicolas; Loskill, Peter; Spengler, Christian; Hümbert, Sebastian; Bischoff, Markus; Jacobs, Karin

    2015-12-01

    The atomic force microscope (AFM) evolved as a standard device in modern microbiological research. However, its capability as a sophisticated force sensor is not used to its full capacity. The AFM turns into a unique tool for quantitative adhesion research in bacteriology by using "bacterial probes". Thereby, bacterial probes are AFM cantilevers that provide a single bacterium or a cluster of bacteria as the contact-forming object. We present a step-by-step protocol for preparing bacterial probes, performing force spectroscopy experiments and processing force spectroscopy data. Additionally, we provide a general insight into the field of bacterial cell force spectroscopy.

  15. A detailed guideline for the fabrication of single bacterial probes used for atomic force spectroscopy.

    PubMed

    Thewes, Nicolas; Loskill, Peter; Spengler, Christian; Hümbert, Sebastian; Bischoff, Markus; Jacobs, Karin

    2015-12-01

    The atomic force microscope (AFM) evolved as a standard device in modern microbiological research. However, its capability as a sophisticated force sensor is not used to its full capacity. The AFM turns into a unique tool for quantitative adhesion research in bacteriology by using "bacterial probes". Thereby, bacterial probes are AFM cantilevers that provide a single bacterium or a cluster of bacteria as the contact-forming object. We present a step-by-step protocol for preparing bacterial probes, performing force spectroscopy experiments and processing force spectroscopy data. Additionally, we provide a general insight into the field of bacterial cell force spectroscopy. PMID:26701715

  16. FRAME (Force Review Automation Environment): MATLAB-based AFM data processor.

    PubMed

    Partola, Kostyantyn R; Lykotrafitis, George

    2016-05-01

    Data processing of force-displacement curves generated by atomic force microscopes (AFMs) for elastic moduli and unbinding event measurements is very time consuming and susceptible to user error or bias. There is an evident need for consistent, dependable, and easy-to-use AFM data processing software. We have developed an open-source software application, the force review automation environment (or FRAME), that provides users with an intuitive graphical user interface, automating data processing, and tools for expediting manual processing. We did not observe a significant difference between manually processed and automatically processed results from the same data sets.

  17. FRAME (Force Review Automation Environment): MATLAB-based AFM data processor.

    PubMed

    Partola, Kostyantyn R; Lykotrafitis, George

    2016-05-01

    Data processing of force-displacement curves generated by atomic force microscopes (AFMs) for elastic moduli and unbinding event measurements is very time consuming and susceptible to user error or bias. There is an evident need for consistent, dependable, and easy-to-use AFM data processing software. We have developed an open-source software application, the force review automation environment (or FRAME), that provides users with an intuitive graphical user interface, automating data processing, and tools for expediting manual processing. We did not observe a significant difference between manually processed and automatically processed results from the same data sets. PMID:26972765

  18. Taking nanomedicine teaching into practice with atomic force microscopy and force spectroscopy.

    PubMed

    Carvalho, Filomena A; Freitas, Teresa; Santos, Nuno C

    2015-12-01

    Atomic force microscopy (AFM) is a useful and powerful tool to study molecular interactions applied to nanomedicine. The aim of the present study was to implement a hands-on atomic AFM course for graduated biosciences and medical students. The course comprises two distinct practical sessions, where students get in touch with the use of an atomic force microscope by performing AFM scanning images of human blood cells and force spectroscopy measurements of the fibrinogen-platelet interaction. Since the beginning of this course, in 2008, the overall rating by the students was 4.7 (out of 5), meaning a good to excellent evaluation. Students were very enthusiastic and produced high-quality AFM images and force spectroscopy data. The implementation of the hands-on AFM course was a success, giving to the students the opportunity of contact with a technique that has a wide variety of applications on the nanomedicine field. In the near future, nanomedicine will have remarkable implications in medicine regarding the definition, diagnosis, and treatment of different diseases. AFM enables students to observe single molecule interactions, enabling the understanding of molecular mechanisms of different physiological and pathological processes at the nanoscale level. Therefore, the introduction of nanomedicine courses in bioscience and medical school curricula is essential.

  19. Taking nanomedicine teaching into practice with atomic force microscopy and force spectroscopy.

    PubMed

    Carvalho, Filomena A; Freitas, Teresa; Santos, Nuno C

    2015-12-01

    Atomic force microscopy (AFM) is a useful and powerful tool to study molecular interactions applied to nanomedicine. The aim of the present study was to implement a hands-on atomic AFM course for graduated biosciences and medical students. The course comprises two distinct practical sessions, where students get in touch with the use of an atomic force microscope by performing AFM scanning images of human blood cells and force spectroscopy measurements of the fibrinogen-platelet interaction. Since the beginning of this course, in 2008, the overall rating by the students was 4.7 (out of 5), meaning a good to excellent evaluation. Students were very enthusiastic and produced high-quality AFM images and force spectroscopy data. The implementation of the hands-on AFM course was a success, giving to the students the opportunity of contact with a technique that has a wide variety of applications on the nanomedicine field. In the near future, nanomedicine will have remarkable implications in medicine regarding the definition, diagnosis, and treatment of different diseases. AFM enables students to observe single molecule interactions, enabling the understanding of molecular mechanisms of different physiological and pathological processes at the nanoscale level. Therefore, the introduction of nanomedicine courses in bioscience and medical school curricula is essential. PMID:26628660

  20. Strength by atomic force microscopy (AFM): Molecular dynamics of water layer squeezing on magnesium oxide

    NASA Astrophysics Data System (ADS)

    Kendall, K.; Dhir, Aman; Yong, Chin W.

    2010-11-01

    Localised strength testing of materials is often carried out in an atomic force microscope (AFM), as foreseen by Kelly in his book Strong Solids (Clarendon Press, Oxford, 1966). During AFM indentation experiments, contamination can strongly influence the observed strength and theoretical interpretation of the results is a major problem. Here, we use molecular dynamics computer modelling to describe the contact of NaCl and MgO crystal probes onto surfaces, comparable to an AFM experiment. Clean NaCl gave elastic, brittle behaviour in contact simulations at 300 K, whereas MgO was more plastic, leading to increased toughness. This paper also considers the strength of an oxide substrate contaminated by water molecules and tested by indentation with a pyramidal probe of oxide crystal. Recent theory on the effect of liquid contaminant layers on surface strength has been mainly focussed on Lennard Jones (LJ) molecules with some studies on alcohols and water, described by molecular dynamics, which allows the molecules to be squeezed out as the crystal lattice is deformed. In this work, we have focused on water by studying the forces between a magnesium oxide (MgO) atomic force microscope (AFM) probe and an MgO slab. Force versus separation has been plotted as the AFM probe was moved towards and away from the substrate. Simulation results showed that the water layers could be removed in steps, giving up to four force peaks. The last monolayer of water could not be squeezed out, even at pressures where MgO deformed plastically. Interestingly, with water present, strength was reduced, but more in tensile than compressive measurements. In conclusion, water contaminating the oxide surface in AFM strength testing is structured. Water layer squeezing removal can be predicted by molecular modelling, which may be verified by AFM experiments to show that water can influence the strength of perfect crystals at the nanometre scale.

  1. Using AFM Force Curves to Explore Properties of Elastomers

    ERIC Educational Resources Information Center

    Ferguson, Megan A.; Kozlowski, Joseph J.

    2013-01-01

    polydimethylsiloxane (PDMS) elastomers. Force curves are used to quantify the stiffness of elastomers prepared with different base-to-curing agent ratios. Trends in observed spring constants of the…

  2. Adhesion of B. subtilis spores and vegetative cells onto stainless steel--DLVO theories and AFM spectroscopy.

    PubMed

    Harimawan, Ardiyan; Zhong, Shaoping; Lim, Chwee-Teck; Ting, Yen-Peng

    2013-09-01

    Interactions between the bacterium Bacillus subtilis (either as vegetative cells or as spores) and stainless steel 316 (SS-316) surfaces were quantified using the classical Derjaguin-Landau-Verwey-Overbeek (DLVO) theory and extended DLVO (xDLVO) approach in conjunction with live force spectroscopy using an Atomic Force Microscope (AFM). The xDLVO approach accounts for acid-base (polar) interactions that are not considered in the classical DLVO theory. AFM results revealed that spores manifested stronger attraction interactions to stainless steel compared to their vegetative cells counterparts due to lower energy barrier as predicted by both the theoretical approaches as well as the higher hydrophobicity on the spore surfaces. Both DLVO and xDLVO theories predict that vegetative cells manifest weaker attachment on the surfaces compared to spores. Results of AFM force measurement corroborate these findings; spores recorded significantly higher adhesion force (2.92±0.4 nN) compared to vegetative cells (0.65±0.2 nN). The adhesion of spores presents greater challenges in biofilm control owing to its stronger attachment and persistence when the spores are formed under adverse environmental conditions. PMID:23777862

  3. Adhesion of B. subtilis spores and vegetative cells onto stainless steel--DLVO theories and AFM spectroscopy.

    PubMed

    Harimawan, Ardiyan; Zhong, Shaoping; Lim, Chwee-Teck; Ting, Yen-Peng

    2013-09-01

    Interactions between the bacterium Bacillus subtilis (either as vegetative cells or as spores) and stainless steel 316 (SS-316) surfaces were quantified using the classical Derjaguin-Landau-Verwey-Overbeek (DLVO) theory and extended DLVO (xDLVO) approach in conjunction with live force spectroscopy using an Atomic Force Microscope (AFM). The xDLVO approach accounts for acid-base (polar) interactions that are not considered in the classical DLVO theory. AFM results revealed that spores manifested stronger attraction interactions to stainless steel compared to their vegetative cells counterparts due to lower energy barrier as predicted by both the theoretical approaches as well as the higher hydrophobicity on the spore surfaces. Both DLVO and xDLVO theories predict that vegetative cells manifest weaker attachment on the surfaces compared to spores. Results of AFM force measurement corroborate these findings; spores recorded significantly higher adhesion force (2.92±0.4 nN) compared to vegetative cells (0.65±0.2 nN). The adhesion of spores presents greater challenges in biofilm control owing to its stronger attachment and persistence when the spores are formed under adverse environmental conditions.

  4. Imaging and force measurement of LDL and HDL by AFM in air and liquid

    PubMed Central

    Gan, Chaoye; Ao, Meiying; Liu, Zhanghua; Chen, Yong

    2015-01-01

    The size and biomechanical properties of lipoproteins are tightly correlated with their structures/functions. While atomic force microscopy (AFM) has been used to image lipoproteins the force measurement of these nano-sized particles is missing. We detected that the sizes of LDL and HDL in liquid are close to the commonly known values. The Young’s modulus of LDL or HDL is ∼0.4 GPa which is similar to that of some viral capsids or nanovesicles but greatly larger than that of various liposomes. The adhesive force of LDL or HDL is small (∼200 pN). The comparison of AFM detection in air and liquid was also performed which is currently lacking. Our data may provide useful information for better understanding and AFM detection of lipoproteins. PMID:25893163

  5. Dynamic force microscopy simulator (dForce): A tool for planning and understanding tapping and bimodal AFM experiments

    PubMed Central

    Guzman, Horacio V; Garcia, Pablo D

    2015-01-01

    Summary We present a simulation environment, dForce, which can be used for a better understanding of dynamic force microscopy experiments. The simulator presents the cantilever–tip dynamics for two dynamic AFM methods, tapping mode AFM and bimodal AFM. It can be applied for a wide variety of experimental situations in air or liquid. The code provides all the variables and parameters relevant in those modes, for example, the instantaneous deflection and tip–surface force, velocity, virial, dissipated energy, sample deformation and peak force as a function of time or distance. The simulator includes a variety of interactions and contact mechanics models to describe AFM experiments including: van der Waals, Hertz, DMT, JKR, bottom effect cone correction, linear viscoelastic forces or the standard linear solid viscoelastic model. We have compared two numerical integration methods to select the one that offers optimal accuracy and speed. The graphical user interface has been designed to facilitate the navigation of non-experts in simulations. Finally, the accuracy of dForce has been tested against numerical simulations performed during the last 18 years. PMID:25821676

  6. Molecular shape and binding force of Mycoplasma mobile's leg protein Gli349 revealed by an AFM study

    SciTech Connect

    Lesoil, Charles; Nonaka, Takahiro; Sekiguchi, Hiroshi; Osada, Toshiya; Miyata, Makoto; Afrin, Rehana; Ikai, Atsushi

    2010-01-15

    Recent studies of the gliding bacteria Mycoplasma mobile have identified a family of proteins called the Gli family which was considered to be involved in this novel and yet fairly unknown motility system. The 349 kDa protein called Gli349 was successfully isolated and purified from the bacteria, and electron microscopy imaging and antibody experiments led to the hypothesis that it acts as the 'leg' of M. mobile, responsible for attachment to the substrate as well as for gliding motility. However, more precise evidence of the molecular shape and function of this protein was required to asses this theory any further. In this study, an atomic force microscope (AFM) was used both as an imaging and a force measurement device to provide new information about Gli349 and its role in gliding motility. AFM images of the protein were obtained revealing a complex structure with both rigid and flexible parts, consistent with previous electron micrographs of the protein. Single-molecular force spectroscopy experiments were also performed, revealing that Gli349 is able to specifically bind to sialyllactose molecules and withstand unbinding forces around 70 pN. These findings strongly support the idea that Gli349 is the 'leg' protein of M. mobile, responsible for binding and also most probably force generation during gliding motility.

  7. Analysis of time-resolved interaction force mode AFM imaging using active and passive probes.

    PubMed

    Giray Oral, Hasan; Parlak, Zehra; Levent Degertekin, F

    2012-09-01

    We present an in-depth analysis of time-resolved interaction force (TRIF) mode imaging for atomic force microscopy (AFM). A nonlinear model of an active AFM probe, performing simultaneous topography and material property imaging on samples with varying elasticity and adhesion is implemented in Simulink®. The model is capable of simulating various imaging modes, probe structures, sample material properties, tip-sample interaction force models, and actuation and feedback schemes. For passive AFM cantilevers, the model is verified by comparing results from the literature. As an example of an active probe, the force sensing integrated readout and active tip (FIRAT) probe is used. Simulation results indicate that the active and damped nature of FIRAT provides a significant level of control over the force applied to the sample, minimizing sample indentation and topography error. Active tip control (ATC) preserves constant contact time during force control for stable contact while preventing the loss of material property information such as elasticity and adhesive force. Simulation results are verified by TRIF mode imaging of the samples with both soft and stiff regions. PMID:22813887

  8. Detection of viruses: atomic force microscopy and surface enhanced raman spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This paper demonstrated the capability of atomic force microscopy (AFM) and surface enhanced Raman spectroscopy (SERS) to function effectively as ultra-sensitive readout tools for chip-scale platforms designed for pathogen detection in complex biological media. AFM allows direct (i.e. label-free) vi...

  9. Quantitative atomic resolution force imaging on epitaxial graphene with reactive and nonreactive AFM probes.

    PubMed

    Boneschanscher, Mark P; van der Lit, Joost; Sun, Zhixiang; Swart, Ingmar; Liljeroth, Peter; Vanmaekelbergh, Daniël

    2012-11-27

    Atomic force microscopy (AFM) images of graphene and graphite show contrast with atomic periodicity. However, the contrast patterns vary depending on the atomic termination of the AFM tip apex and the tip-sample distance, hampering the identification of the atomic positions. Here, we report quantitative AFM imaging of epitaxial graphene using inert (carbon-monoxide-terminated) and reactive (iridium-terminated) tips. The atomic image contrast is markedly different with these tip terminations. With a reactive tip, we observe an inversion from attractive to repulsive atomic contrast with decreasing tip-sample distance, while a nonreactive tip only yields repulsive atomic contrast. We are able to identify the atoms with both tips at any tip-sample distance. This is a prerequisite for future structural and chemical analysis of adatoms, defects, and the edges of graphene nanostructures, crucial for understanding nanoscale graphene devices.

  10. Force-controlled manipulation of single cells: from AFM to FluidFM.

    PubMed

    Guillaume-Gentil, Orane; Potthoff, Eva; Ossola, Dario; Franz, Clemens M; Zambelli, Tomaso; Vorholt, Julia A

    2014-07-01

    The ability to perturb individual cells and to obtain information at the single-cell level is of central importance for addressing numerous biological questions. Atomic force microscopy (AFM) offers great potential for this prospering field. Traditionally used as an imaging tool, more recent developments have extended the variety of cell-manipulation protocols. Fluidic force microscopy (FluidFM) combines AFM with microfluidics via microchanneled cantilevers with nano-sized apertures. The crucial element of the technology is the connection of the hollow cantilevers to a pressure controller, allowing their operation in liquid as force-controlled nanopipettes under optical control. Proof-of-concept studies demonstrated a broad spectrum of single-cell applications including isolation, deposition, adhesion and injection in a range of biological systems. PMID:24856959

  11. PREFACE: NC-AFM 2004: Proceedings of the 7th International Conference on Non-contact Atomic Force Microscopy

    NASA Astrophysics Data System (ADS)

    Schwarz, Udo

    2005-03-01

    With the ongoing miniaturization of devices and controlled nanostructuring of materials, the importance of atomic-scale information on surfaces and surface properties is growing continuously. The astonishing progress in nanoscience and nanotechnology that took place during the last two decades was in many ways related to recent progress in high-resolution imaging techniques such as scanning tunnelling microscopy and transmission electron microscopy. Since the mid-1990s, non-contact atomic force microscopy (NC-AFM) performed in ultrahigh vacuum has evolved as an alternative technique that achieves atomic resolution, but without the restriction to conducting surfaces of the previously established techniques. Advances of the rapidly developing field of NC-AFM are discussed at annual conferences as part of a series that started in 1998 in Osaka, Japan. This special issue of Nanotechnology is a compilation of original work presented at the 7th International Conference on Non-contact Atomic Force Microscopy that took place in Seattle, USA, 12-15 September 2004. Over the years, the conference grew in size and scope. Atomic resolution imaging of oxides and semiconductors remains an issue. Noticeable new developments have been presented in this regard such as, e.g., the demonstrated ability to manipulate individual atoms. Additionally, the investigation of individual molecules, clusters, and organic materials gains more and more attention. In this context, considerable effort is undertaken to transfer the NC-AFM principle based on frequency modulation to applications in air and liquids with the goal of enabling high-resolution surface studies of biological material in native environments, as well as to reduce the experimental complexity, which so far involves the availability of (costly) vacuum systems. Force spectroscopy methods continue to be improved and are applied to topics such as the imaging of the three-dimensional force field as a function of the distance with

  12. Radiation pressure excitation of Low Temperature Atomic Force & Magnetic Force Microscope (LT-AFM/MFM) for Imaging

    NASA Astrophysics Data System (ADS)

    Karci, Ozgur; Celik, Umit; Oral, Ahmet; NanoMagnetics Instruments Ltd. Team; Middle East Tech Univ Team

    2015-03-01

    We describe a novel method for excitation of Atomic Force Microscope (AFM) cantilevers by means of radiation pressure for imaging in an AFM for the first time. Piezo excitation is the most common method for cantilever excitation, but it may cause spurious resonance peaks. A fiber optic interferometer with 1310 nm laser was used both to measure the deflection of cantilever and apply a force to the cantilever in a LT-AFM/MFM from NanoMagnetics Instruments. The laser power was modulated at the cantilever`s resonance frequency by a digital Phase Lock Loop (PLL). The force exerted by the radiation pressure on a perfectly reflecting surface by a laser beam of power P is F = 2P/c. We typically modulate the laser beam by ~ 800 μW and obtain 10nm oscillation amplitude with Q ~ 8,000 at 2.5x10-4 mbar. The cantilever's stiffness can be accurately calibrated by using the radiation pressure. We have demonstrated performance of the radiation pressure excitation in AFM/MFM by imaging a hard disk sample between 4-300K and Abrikosov vortex lattice in BSCCO single crystal at 4K to for the first time.

  13. Dynamics of a disturbed sessile drop measured by atomic force microscopy (AFM).

    PubMed

    McGuiggan, Patricia M; Grave, Daniel A; Wallace, Jay S; Cheng, Shengfeng; Prosperetti, Andrea; Robbins, Mark O

    2011-10-01

    A new method for studying the dynamics of a sessile drop by atomic force microscopy (AFM) is demonstrated. A hydrophobic microsphere (radius, r ∼ 20-30 μm) is brought into contact with a small sessile water drop resting on a polytetrafluoroethylene (PTFE) surface. When the microsphere touches the liquid surface, the meniscus rises onto it because of capillary forces. Although the microsphere volume is 6 orders of magnitude smaller than the drop, it excites the normal resonance modes of the liquid interface. The sphere is pinned at the interface, whose small (<100 nm) oscillations are readily measured with AFM. Resonance oscillation frequencies were measured for drop volumes between 5 and 200 μL. The results for the two lowest normal modes are quantitatively consistent with continuum calculations for the natural frequency of hemispherical drops with no adjustable parameters. The method may enable sensitive measurements of volume, surface tension, and viscosity of small drops.

  14. Nanoscopic imaging of mechanical properties of metal films with magnetic-force-controlled AFM

    NASA Astrophysics Data System (ADS)

    Yamamoto, S.-I.; Yamada, H.; Matsushige, K.; Ishida, T.; Mizutani, W.; Tokumoto, H.

    1999-08-01

    We have succeeded in measuring the Young's moduli of inorganic films with a diamond tip of 160-200 nm in diameter using magnetic-force-controlled atomic force microscopy (MFC-AFM). The measurement of Young's moduli was carried out by detecting the tip amplitude on the sample with an applied force of 20 nN. This technique was applied successfully to obtain Young's moduli mapping of gold islands with 1 nm thickness and nickel films with 10 nm thickness. We obtained an effective spring constant of 3000 N m -1 on gold and 21 000 N m -1 on nickel using MFC-AFM. We could normalize the effective area of the contact radius between the tip and the sample. A value of 350 GPa was derived for the Young's modulus of the nickel film, indicative of the elasticity of native nickel oxide (NiO 2). This shows that the MFC-AFM technique measures the mechanical properties of a thin layer on top of the sample.

  15. Direct measurement of optical force induced by near-field plasmonic cavity using dynamic mode AFM

    DOE PAGES

    Guan, Dongshi; Hang, Zhi Hong; Marset, Zsolt; Liu, Hui; Kravchenko, Ivan I.; Chan, Ho Bun; Chan, C. T.; Tong, Penger

    2015-11-20

    Plasmonic nanostructures have attracted much attention in recent years because of their potential applications in optical manipulation through near-field enhancement. Continuing experimental efforts have been made to develop accurate techniques to directly measure the near-field optical force induced by the plasmonic nanostructures in the visible frequency range. In this work, we report a new application of dynamic mode atomic force microscopy (DM-AFM) in the measurement of the enhanced optical force acting on a nano-structured plasmonic resonant cavity. The plasmonic cavity is made of an upper gold-coated glass sphere and a lower quartz substrate patterned with an array of subwavelength goldmore » disks. In the near-field when the sphere is positioned close to the disk array, plasmonic resonance is excited in the cavity and the induced force by a 1550 nm infrared laser is found to be increased by an order of magnitude compared with the photon pressure generated by the same laser light. Lastly, the experiment demonstrates that DM-AFM is a powerful tool for the study of light induced forces and their enhancement in plasmonic nanostructures.« less

  16. Direct measurement of optical force induced by near-field plasmonic cavity using dynamic mode AFM

    SciTech Connect

    Guan, Dongshi; Hang, Zhi Hong; Marset, Zsolt; Liu, Hui; Kravchenko, Ivan I.; Chan, Ho Bun; Chan, C. T.; Tong, Penger

    2015-11-20

    Plasmonic nanostructures have attracted much attention in recent years because of their potential applications in optical manipulation through near-field enhancement. Continuing experimental efforts have been made to develop accurate techniques to directly measure the near-field optical force induced by the plasmonic nanostructures in the visible frequency range. In this work, we report a new application of dynamic mode atomic force microscopy (DM-AFM) in the measurement of the enhanced optical force acting on a nano-structured plasmonic resonant cavity. The plasmonic cavity is made of an upper gold-coated glass sphere and a lower quartz substrate patterned with an array of subwavelength gold disks. In the near-field when the sphere is positioned close to the disk array, plasmonic resonance is excited in the cavity and the induced force by a 1550 nm infrared laser is found to be increased by an order of magnitude compared with the photon pressure generated by the same laser light. Lastly, the experiment demonstrates that DM-AFM is a powerful tool for the study of light induced forces and their enhancement in plasmonic nanostructures.

  17. Direct Measurement of Optical Force Induced by Near-Field Plasmonic Cavity Using Dynamic Mode AFM

    PubMed Central

    Guan, Dongshi; Hang, Zhi Hong; Marcet, Zsolt; Liu, Hui; Kravchenko, I. I.; Chan, C. T.; Chan, H. B.; Tong, Penger

    2015-01-01

    Plasmonic nanostructures have attracted much attention in recent years because of their potential applications in optical manipulation through near-field enhancement. Continuing experimental efforts have been made to develop accurate techniques to directly measure the near-field optical force induced by the plasmonic nanostructures in the visible frequency range. In this work, we report a new application of dynamic mode atomic force microscopy (DM-AFM) in the measurement of the enhanced optical force acting on a nano-structured plasmonic resonant cavity. The plasmonic cavity is made of an upper gold-coated glass sphere and a lower quartz substrate patterned with an array of subwavelength gold disks. In the near-field when the sphere is positioned close to the disk array, plasmonic resonance is excited in the cavity and the induced force by a 1550 nm infrared laser is found to be increased by an order of magnitude compared with the photon pressure generated by the same laser light. The experiment demonstrates that DM-AFM is a powerful tool for the study of light induced forces and their enhancement in plasmonic nanostructures. PMID:26586455

  18. Characterization and Optimization of Quartz Tuning Fork-Based Force Sensors for Combined STM/AFM

    NASA Astrophysics Data System (ADS)

    Castellanos-Gomez, Andres; Agraït, Nicolás; Rubio-Bollinger, Gabino

    This chapter will be divided in two main parts. In the first one, we will show a detailed analysis of the dynamics of quartz tuning fork resonators which are being increasingly used in scanning probe microscopy as force sensors. We will also show that a coupled harmonic oscillators model, which includes a finite coupling between the prongs, is in remarkable agreement with the observed motion of the tuning forks. Relevant parameters for the tuning fork performance such as the effective spring constant can be obtained from our analysis. In the second one, we will present an implementation of a quartz tuning fork supplemented with optimized tips based on carbon fibers. The remarkable electrical and mechanical properties of carbon fiber make these tips more suitable for combined and/or simultaneous STM and AFM than conventional metallic tips. The fabrication and the characterization of these carbon fiber tips as well as their performance in STM/AFM will be detailed.

  19. Molecular Force Spectroscopy on Cells

    NASA Astrophysics Data System (ADS)

    Liu, Baoyu; Chen, Wei; Zhu, Cheng

    2015-04-01

    Molecular force spectroscopy has become a powerful tool to study how mechanics regulates biology, especially the mechanical regulation of molecular interactions and its impact on cellular functions. This force-driven methodology has uncovered a wealth of new information of the physical chemistry of molecular bonds for various biological systems. The new concepts, qualitative and quantitative measures describing bond behavior under force, and structural bases underlying these phenomena have substantially advanced our fundamental understanding of the inner workings of biological systems from the nanoscale (molecule) to the microscale (cell), elucidated basic molecular mechanisms of a wide range of important biological processes, and provided opportunities for engineering applications. Here, we review major force spectroscopic assays, conceptual developments of mechanically regulated kinetics of molecular interactions, and their biological relevance. We also present current challenges and highlight future directions.

  20. Spatially resolved force spectroscopy of bacterial surfaces using force-volume imaging.

    PubMed

    Gaboriaud, Fabien; Parcha, Bhargava S; Gee, Michelle L; Holden, James A; Strugnell, Richard A

    2008-04-01

    Force spectroscopy using the atomic force microscope (AFM) is a powerful technique for measuring physical properties and interaction forces at microbial cell surfaces. Typically for such a study, the point at which a force measurement will be made is located by first imaging the cell using AFM in contact mode. In this study, we image the bacterial cell Shewanella putrefaciens for subsequent force measurements using AFM in force-volume mode and compare this to contact-mode images. It is known that contact-mode imaging does not accurately locate the apical surface and periphery of the cell since, in contact mode, a component of the applied load laterally deforms the cell during the raster scan. Here, we illustrate that contact-mode imaging does not accurately locate the apical surface and periphery of the cell since, in contact mode, a component of the applied load laterally deforms the cell during the raster scan. This is an artifact due to the deformability and high degree of curvature of bacterial cells. We further show that force-volume mode imaging avoids the artifacts associated with contact-mode imaging due to surface deformation since it involves the measurement of a grid of individual force profiles. The topographic image is subsequently reconstructed from the zero-force height (the contact distance between the AFM tip and the surface) at each point on the cell surface. We also show how force-volume measurements yield applied load versus indentation data from which mechanical properties of the cell such as Young's modulus, cell turgor pressure and elastic and plastic energies can be extracted.

  1. Interaction force measurement between E. coli cells and nanoparticles immobilized surfaces by using AFM

    SciTech Connect

    Zhang, Wen; Chen, Yongsheng

    2011-01-01

    To better understand environmental behaviors of nanoparticles (NPs), we used the atomic force microscopy (AFM) to measure interaction forces between E. coli cells and NPs immobilized on surfaces in an aqueous environment. The results showed that adhesion force strength was significantly influenced by particle size for both hematite ( -Fe2 O3 ) and corundum ( -Al2 O3 ) NPs whereas the effect on the repulsive force was not observed. The adhesion force decreased from 6.3 0.7 nN to 0.8 0.4 nN as hematite NPs increased from 26 nm to 98 nm in diameter. Corundum NPs exhibited a similar dependence of adhesion force on particle size. The Johnson Kendall Roberts (JKR) model was employed to estimate the contact area between E. coli cells and NPs, and based on the JKR model a new model that considers local effective contact area was developed. The prediction of the new model matched the size dependence of adhesion force in experimental results. Size effects on adhesion forces may originate from the difference in local effective contact areas as supported by our model. These findings provide fundamental information for interpreting the environmental behaviors and biological interactions of NPs, which barely have been addressed.

  2. Oscillatory structural forces due to nonionic surfactant micelles: data by colloidal-probe AFM vs theory.

    PubMed

    Christov, Nikolay C; Danov, Krassimir D; Zeng, Yan; Kralchevsky, Peter A; von Klitzing, Regine

    2010-01-19

    Micellar solutions of nonionic surfactants Brij 35 and Tween 20 are confined between two surfaces in a colloidal-probe atomic-force microscope (CP-AFM). The experimentally detected oscillatory forces due to the layer-by-layer expulsion of the micelles agree very well with the theoretical predictions for hard-sphere fluids. While the experiment gives parts of the stable branches of the force curve, the theoretical model allows reconstruction of the full oscillatory curve. Therewith, the strength and range of the ordering could be determined. The resulting aggregation number from the fits of the force curves for Brij 35 is close to 70 and exhibits a slight tendency to increase with the surfactant concentration. The last layer of micelles cannot be pressed out. The measured force-vs-distance curve has nonequilibrium portions, which represent "jumps" from one to another branch of the respective equilibrium oscillatory curve. In the case of Brij 35, at concentrations <150 mM spherical micelles are present and the oscillation period is close to the micelle diameter, slightly decreasing with the rise of concentration. For elongated micelles (at concentration 200 mM), no harmonic oscillations are observed anymore; instead, the period increases with the decrease of film thickness. In the case of Tween 20, the force oscillations are almost suppressed, which implies that the micelles of this surfactant are labile and are demolished by the hydrodynamic shear stresses due to the colloidal-probe motion. The comparison of the results for the two surfactants demonstrates that in some cases the micelles can be destroyed by the CP-AFM, but in other cases they can be stable and behave as rigid particles. This behavior correlates with the characteristic times of the slow micellar relaxation process for these surfactants.

  3. 3D force and displacement sensor for SFA and AFM measurements.

    PubMed

    Kristiansen, Kai; McGuiggan, Patricia; Carver, Greg; Meinhart, Carl; Israelachvili, Jacob

    2008-02-19

    A new device has been designed, and a prototype built and tested, that can simultaneously measure the displacements and/or the components of a force in three orthogonal directions. The "3D sensor" consists of four or eight strain gauges attached to the four arms of a single cross-shaped force-measuring cantilever spring. Finite element modeling (FEM) was performed to optimize the design configuration to give desired sensitivity of force, displacement, stiffness, and resonant frequency in each direction (x, y, and z) which were tested on a "mesoscale" device and found to agree with the predicted values to within 4-10%. The device can be fitted into a surface forces apparatus (SFA), and a future smaller "microscale" microfabricated version can be fitted into an atomic force microscope (AFM) for simultaneous measurements of the normal and lateral (friction) forces between a tip (or colloidal bead probe) and a surface, and the topography of the surface. Results of the FEM analysis are presented, and approximate equations derived using linear elasticity theory are given for the sensitivity in each direction. Initial calibrations and measurements of thin film rheology (lubrication forces) using the "mesoscale" prototype show the device to function as expected.

  4. Following aptamer-ricin specific binding by single molecule recognition and force spectroscopy measurements

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The atomic force microscope (AFM) recognition and dynamic force spectroscopy (DFS) experiments provide both morphology and interaction information of the aptamer and protein, which can be used for the future study on the thermodynamics and kinetics properties of ricin-aptamer/antibody interactions. ...

  5. Probing correlated current and force effects of nanoparticle charge states by hybrid STM-AFM

    NASA Astrophysics Data System (ADS)

    Suganuma, Y.; Trudeau, P.-E.; Dhirani, A.-A.

    2002-12-01

    By using an atomic force microscope (AFM) cantilever as a substrate for a scanning tunneling microscope (STM), we can simultaneously probe currents I and forces F in a tunnel junction as a function of bias voltage V. Measurements performed using gold-nanoparticle (NP) coated cantilevers under ambient conditions reveal correlated kinks in both I-V and F-V curves. Changes in background charge distributions, which have been problematic in controlling single charges in such systems, can be readily detected by the hybrid STM-AFM. To test whether we can statistically attribute observed kinks to Coulomb staircase phenomena, we have performed measurements using NP’s of different sizes. NP’s 4.8 and 2.5 nm in diameter exhibit kinks spaced on average 0.22 and 0.52 V apart, respectively. These values are in good agreement with a metallic sphere flat surface model for the STM tip NP capacitance and the orthodox model for single-electron tunneling.

  6. Diamond-modified AFM probes: from diamond nanowires to atomic force microscopy-integrated boron-doped diamond electrodes.

    PubMed

    Smirnov, Waldemar; Kriele, Armin; Hoffmann, René; Sillero, Eugenio; Hees, Jakob; Williams, Oliver A; Yang, Nianjun; Kranz, Christine; Nebel, Christoph E

    2011-06-15

    In atomic force microscopy (AFM), sharp and wear-resistant tips are a critical issue. Regarding scanning electrochemical microscopy (SECM), electrodes are required to be mechanically and chemically stable. Diamond is the perfect candidate for both AFM probes as well as for electrode materials if doped, due to diamond's unrivaled mechanical, chemical, and electrochemical properties. In this study, standard AFM tips were overgrown with typically 300 nm thick nanocrystalline diamond (NCD) layers and modified to obtain ultra sharp diamond nanowire-based AFM probes and probes that were used for combined AFM-SECM measurements based on integrated boron-doped conductive diamond electrodes. Analysis of the resonance properties of the diamond overgrown AFM cantilevers showed increasing resonance frequencies with increasing diamond coating thicknesses (i.e., from 160 to 260 kHz). The measured data were compared to performed simulations and show excellent correlation. A strong enhancement of the quality factor upon overgrowth was also observed (120 to 710). AFM tips with integrated diamond nanowires are shown to have apex radii as small as 5 nm and where fabricated by selectively etching diamond in a plasma etching process using self-organized metal nanomasks. These scanning tips showed superior imaging performance as compared to standard Si-tips or commercially available diamond-coated tips. The high imaging resolution and low tip wear are demonstrated using tapping and contact mode AFM measurements by imaging ultra hard substrates and DNA. Furthermore, AFM probes were coated with conductive boron-doped and insulating diamond layers to achieve bifunctional AFM-SECM probes. For this, focused ion beam (FIB) technology was used to expose the boron-doped diamond as a recessed electrode near the apex of the scanning tip. Such a modified probe was used to perform proof-of-concept AFM-SECM measurements. The results show that high-quality diamond probes can be fabricated, which are

  7. Study of Schottky contact between Au and NiO nanowire by conductive atomic force microscopy (C-AFM): The case of surface states

    NASA Astrophysics Data System (ADS)

    Zhang, Yidong

    2015-05-01

    In this work, NiO nanowires have been synthesized by a hydrothermal reaction of NiCl2 with Na2C2O4 in the presence of ethylene glycol at 180 °C for 12 h, then calcinated at 400 °C for 2 h. The NiO nanowires were analyzed by means of scanning electron microscope (SEM), atomic force microscope (AFM), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS). The resulting current-voltage (I-V) characteristics of the NiO nanowires exhibited a clear rectifying behavior. This rectify behavior was attributed to the formation of a Schottky contact between Au coated atomic force microscopy (AFM) tip and NiO nanowires (nano-M/SC) which was dominated by the surface states in NiO itself. Photo-assisted conductive AFM (PC-AFM) was used to demonstrate how the I-V characteristics are influenced by the surface states. Our I-V results also showed that the nano-M/SCs had a good photoelectric switching effect at reverse bias.

  8. Medical applications of atomic force microscopy and Raman spectroscopy.

    PubMed

    Choi, Samjin; Jung, Gyeong Bok; Kim, Kyung Sook; Lee, Gi-Ja; Park, Hun-Kuk

    2014-01-01

    This paper reviews the recent research and application of atomic force microscopy (AFM) and Raman spectroscopy techniques, which are considered the multi-functional and powerful toolkits for probing the nanostructural, biomechanical and physicochemical properties of biomedical samples in medical science. We introduce briefly the basic principles of AFM and Raman spectroscopy, followed by diagnostic assessments of some selected diseases in biomedical applications using them, including mitochondria isolated from normal and ischemic hearts, hair fibers, individual cells, and human cortical bone. Finally, AFM and Raman spectroscopy applications to investigate the effects of pharmacotherapy, surgery, and medical device therapy in various medicines from cells to soft and hard tissues are discussed, including pharmacotherapy--paclitaxel on Ishikawa and HeLa cells, telmisartan on angiotensin II, mitomycin C on strabismus surgery and eye whitening surgery, and fluoride on primary teeth--and medical device therapy--collagen cross-linking treatment for the management of progressive keratoconus, radiofrequency treatment for skin rejuvenation, physical extracorporeal shockwave therapy for healing of Achilles tendinitis, orthodontic treatment, and toothbrushing time to minimize the loss of teeth after exposure to acidic drinks.

  9. Colloid-probe AFM studies of the interaction forces of proteins adsorbed on colloidal crystals.

    PubMed

    Singh, Gurvinder; Bremmell, Kristen E; Griesser, Hans J; Kingshott, Peter

    2015-04-28

    In recent years, colloid-probe AFM has been used to measure the direct interaction forces between colloidal particles of different size or surface functionality in aqueous media, as one can study different forces in symmerical systems (i.e., sphere-sphere geometry). The present study investigates the interaction between protein coatings on colloid probes and hydrophilic surfaces decorated with hexagonally close packed single particle layers that are either uncoated or coated with proteins. Controlled solvent evaporation from aqueous suspensions of colloidal particles (coated with or without lysozyme and albumin) produces single layers of close-packed colloidal crystals over large areas on a solid support. The measurements have been carried out in an aqueous medium at different salt concentrations and pH values. The results show changes in the interaction forces as the surface charge of the unmodified or modified particles, and ionic strength or pH of the solution is altered. At high ionic strength or pH, electrostatic interactions are screened, and a strong repulsive force at short separation below 5 nm dominates, suggesting structural changes in the absorbed protein layer on the particles. We also study the force of adhesion, which decreases with an increment in the salt concentration, and the interaction between two different proteins indicating a repulsive interaction on approach and adhesion on retraction. PMID:25758979

  10. Forced Unfolding of the Coiled-Coils of Fibrinogen by Single-Molecule AFM

    NASA Astrophysics Data System (ADS)

    Brown, Andre; Litvinov, Rustem; Discher, Dennis; Weisel, John

    2007-03-01

    A blood clot needs to have the right degree of stiffness and plasticity for hemostasis, but the origin of these mechanical properties is unknown. Here we report the first measurements using single molecule atomic force microscopy (AFM) to study the forced unfolding of fibrinogen to begin addressing this problem. To generate longer reproducible curves than are possible using monomer, factor XIIIa cross-linked, single chain fibrinogen oligomers were used. When extended under force, these oligomers showed sawtooth shaped force-extension patterns characteristic of unfolding proteins with a peak-to-peak separation of approximately 26 nm, consistent with the independent unfolding of the coiled-coils. These results were then reproduced using a Monte Carlo simulation with parameters in the same range as those previously used for unfolding globular domains. In particular, we found that the refolding time was negligible on experimental time and force scales in contrast to previous work on simpler coiled-coils. We suggest that this difference may be due to fibrinogen's structurally and topologically more complex coiled-coils and that an interaction between the alpha C and central domains may be involved. These results suggest a new functional property of fibrinogen and that the coiled-coil is more than a passive structural element of this molecule.

  11. Effect of enamel morphology on nanoscale adhesion forces of streptococcal bacteria : An AFM study.

    PubMed

    Wang, Chuanyong; Zhao, Yongqi; Zheng, Sainan; Xue, Jing; Zhou, Jinglin; Tang, Yi; Jiang, Li; Li, Wei

    2015-01-01

    We explore the influence of enamel surface morphology on nanoscale bacterial adhesion forces. Three dimensional morphology characteristics of enamel slices, which were treated with phosphoric acid (for 0 s, 5 s, 10 s, 20 s, and 30 s), were acquired. Adhesion forces of three initial colonizers (Streptococcus oralis, Streptococcus sanguinis, and Streptococcus mitis) and two cariogenic bacterial strains (Streptococcus mutans and Streptococcus sobrinus) with etched enamel surfaces were determined. Comparison of the forces was made by using bacterial probe method under atomic force microscope (AFM) in adhesion buffer. The results showed that enamel morphology was significantly altered by etching treatment. The roughness, peak-to-valley height, and valley-to-valley width of the depth profile, surface area, and volume increased linearly with acid exposure time, and reached the maximum at 30s, respectively. The adhesion forces of different strains increased accordingly with etching time. Adhesion forces of S. oralis, S. mitis, S. mutans, and S. sobrinus reached the maximum values of 0.81 nN, 0.84 nN, 0.73 nN, and 0.64 nN with enamel treated for 20s, respectively, whereas that of S. sanguinis at 10s (1.28nN), and dropped on coarser enamel surfaces. In conclusion, enamel micro-scale morphology may significantly alter the direct adhesion forces of bacteria. And there may be a threshold roughness for bacterial adhesion on enamel surface.

  12. Scanning electron and atomic force microscopy, and raman and x-ray photoelectron spectroscopy characterization of near-isogenic soft and hard wheat kernels and corresponding flours

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Atomic force microscopy (AFM), Raman spectroscopy and X-ray photoelectron spectroscopy (XPS) are used to investigate vitreous (hard) and non-vitreous (soft) wheat kernels and their corresponding wheat flours. AFM data reveal two different microstructures. The vitreous kernel reveals a granular text...

  13. Nanomechanical characterization of nanostructured bainitic steel: Peak Force Microscopy and Nanoindentation with AFM.

    PubMed

    Morales-Rivas, Lucia; González-Orive, Alejandro; Garcia-Mateo, Carlos; Hernández-Creus, Alberto; Caballero, Francisca G; Vázquez, Luis

    2015-01-01

    The full understanding of the deformation mechanisms in nanostructured bainite requires the local characterization of its mechanical properties, which are expected to change from one phase, bainitic ferrite, to another, austenite. This study becomes a challenging process due to the bainitic nanostructured nature and high Young's modulus. In this work, we have carried out such study by means of the combination of AFM-based techniques, such as nanoindentation and Peak Force Quantitative Nanomechanical Mapping (PF-QNM) measurements. We have addressed critically the limits and advantages of these techniques and been able to measure some elastoplastic parameters of both phases. Specifically, we have analyzed by PF-QNM two nanostructured bainitic steels, with a finer and a coarser structure, and found that both phases have a similar Young's modulus. PMID:26602631

  14. Nanomechanical characterization of nanostructured bainitic steel: Peak Force Microscopy and Nanoindentation with AFM

    PubMed Central

    Morales-Rivas, Lucia; González-Orive, Alejandro; Garcia-Mateo, Carlos; Hernández-Creus, Alberto; Caballero, Francisca G.; Vázquez, Luis

    2015-01-01

    The full understanding of the deformation mechanisms in nanostructured bainite requires the local characterization of its mechanical properties, which are expected to change from one phase, bainitic ferrite, to another, austenite. This study becomes a challenging process due to the bainitic nanostructured nature and high Young’s modulus. In this work, we have carried out such study by means of the combination of AFM-based techniques, such as nanoindentation and Peak Force Quantitative Nanomechanical Mapping (PF-QNM) measurements. We have addressed critically the limits and advantages of these techniques and been able to measure some elastoplastic parameters of both phases. Specifically, we have analyzed by PF-QNM two nanostructured bainitic steels, with a finer and a coarser structure, and found that both phases have a similar Young’s modulus. PMID:26602631

  15. Dynamic force spectroscopy of parallel individual mucin1-antibody bonds

    SciTech Connect

    Sulchek, T A; Friddle, R W; Langry, K; Lau, E; Albrecht, H; Ratto, T; DeNardo, S; Colvin, M E; Noy, A

    2005-05-02

    We used atomic force microscopy (AFM) to measure the binding forces between Mucin1 (MUC1) peptide and a single chain antibody fragment (scFv) selected from a scFv library screened against MUC1. This binding interaction is central to the design of the molecules for targeted delivery of radioimmunotherapeutic agents for prostate and breast cancer treatment. Our experiments separated the specific binding interaction from non-specific interactions by tethering the antibody and MUC1 molecules to the AFM tip and sample surface with flexible polymer spacers. Rupture force magnitude and elastic characteristics of the spacers allowed identification of the bond rupture events corresponding to different number of interacting proteins. We used dynamic force spectroscopy to estimate the intermolecular potential widths and equivalent thermodynamic off rates for mono-, bi-, and tri-valent interactions. Measured interaction potential parameters agree with the results of molecular docking simulation. Our results demonstrate that an increase of the interaction valency leads to a precipitous decline in the dissociation rate. Binding forces measured for mono and multivalent interactions match the predictions of a Markovian model for the strength of multiple uncorrelated bonds in parallel configuration. Our approach is promising for comparison of the specific effects of molecular modifications as well as for determination of the best configuration of antibody-based multivalent targeting agents.

  16. Combined quantitative ultrasonic and time-resolved interaction force AFM imaging

    SciTech Connect

    Parlak, Z.; Degertekin, F. L.

    2011-01-15

    The authors describe a method where quantitative ultrasonic atomic force microscopy (UAFM) is achieved during time-resolved interaction force (TRIF) imaging in intermittent contact mode. The method uses a calibration procedure for quantitative UAFM. It improves elasticity measurements of stiff regions of surfaces while retaining the capabilities of the TRIF mode for topography, adhesion, dissipation, and elasticity measurements on soft regions of sample surfaces. This combination is especially advantageous when measuring and imaging samples with broad stiffness range in a nondestructive manner. The experiments utilize an active AFM probe with high bandwidth and the UAFM calibration is performed by measuring the magnitude of the time-resolved UAFM signal at a judiciously chosen frequency for different contact stiffness values during individual taps. Improved sensitivity to stiff surface elasticity is demonstrated on a special sample. The results show that combining UAFM with TRIF provides 2.5 GPa (5%) standard deviation on the silicon surface reduced Young's modulus, representing 5x improvement over using only TRIF mode imaging.

  17. Accurate force spectroscopy in tapping mode atomic force microscopy in liquids

    NASA Astrophysics Data System (ADS)

    Xu, Xin; Melcher, John; Raman, Arvind

    2010-01-01

    Existing force spectroscopy methods in tapping mode atomic force microscopy (AFM) such as higher harmonic inversion [M. Stark, R. W. Stark, W. M. Heckl, and R. Guckenberger, Proc. Natl. Acad. Sci. U.S.A. 99, 8473 (2002)] or scanning probe acceleration microscopy [J. Legleiter, M. Park, B. Cusick, and T. Kowalewski, Proc. Natl. Acad. Sci. U.S.A. 103, 4813 (2006)] or integral relations [M. Lee and W. Jhe, Phys. Rev. Lett. 97, 036104 (2006); S. Hu and A. Raman, Nanotechnology 19, 375704 (2008); H. Hölscher, Appl. Phys. Lett. 89, 123109 (2006); A. J. Katan, Nanotechnology 20, 165703 (2009)] require and assume as an observable the tip dynamics in a single eigenmode of the oscillating microcantilever. We demonstrate that this assumption can distort significantly the extracted tip-sample interaction forces when applied to tapping mode AFM with soft cantilevers in liquid environments. This exception is due to the fact that under these conditions the second eigenmode is momentarily excited and the observed tip dynamics clearly contains contributions from the fundamental and second eigenmodes. To alleviate this problem, a simple experimental method is proposed to screen the second eigenmode contributions in the observed tip deflection signal to allow accurate tip-sample force reconstruction in liquids. The method is implemented experimentally to reconstruct interaction forces on polymer, bacteriorhodopsin membrane, and mica samples in buffer solutions.

  18. Study of pluronic F68 molecules on silicon with Atomic Force Microscopy (AFM)

    NASA Astrophysics Data System (ADS)

    Retamal, M. J.; Volkmann, U. G.; Samith, V. D.

    2011-03-01

    The triblock copolymer pluronic F68 belongs to a class of amphiphilic nonionic surfactants, relevant for applications in medicine, which includes transport of drugs to selective targets in the human body. In the present work, pluronic F68 films have been deposited on Si/SiO2 substrates. Previous to deposition, Si/SiO2 substrates were cleaned in acid and afterwards rinsed with ultra pure water. This cleaning method leaves intact the silicon oxide layer and provides a hydrophilic surface. The aliquots were prepared from aqueous solutions, obtaining different concentrations, which were deposited on the substrates and dried at room temperature. Each of these dispersions is in the range from 0.5 x 10-4 M to 10.0 x 10-4 M. Atomic force microscopy (AFM) shows changes in the morphology of the films, caused by the gradual increase of concentration. These changes occur in a narrow range of concentrations, attributed to the critical micelle concentration (CMC). Supramolecular structures (clusters) coexist at the CMC and above, forming 3D structures such as ``dendritics.'' The percentage of F68 coverage on the substrate depends on the increase in molar concentration. In a ``coverage vs. molar concentration'' plot we obtain a curve with an inflection point that coincides with the CMC reported for a variety of techniques and conditions.

  19. AFM PeakForce QNM mode: Evidencing nanometre-scale mechanical properties of chitin-silica hybrid nanocomposites.

    PubMed

    Smolyakov, G; Pruvost, S; Cardoso, L; Alonso, B; Belamie, E; Duchet-Rumeau, J

    2016-10-20

    PeakForce Quantitative Nanomechanical Mapping (QNM) AFM mode was used to explore the mechanical properties of textured chitin-silica hybrid films at the nanoscale. The influence of the force applied by the tip on the sample surface was studied for standard homogeneous samples, for chitin nanorods and for chitin-silica hybrid nanocomposites. Thick films of superimposed chitin nanorods showed a monotonous increase of DMT modulus (based on the Derjaguin-Muller-Toporov model) owing to an increase in modulus at the interface between nanorods due to geometrical constraints of the AFM acquisition. A similar variation of DMT modulus was obtained for chitin-silica hybrid thick films related to mechanical strengthening induced by the presence of silica. This work revealed the role of the organic-inorganic interface, at the nanoscale, in the mechanical behaviour of textured materials using PeakForce QNM mode, with optimized analysis conditions. PMID:27474579

  20. High-resolution atomic force microscopy and spectroscopy of native membrane proteins

    NASA Astrophysics Data System (ADS)

    Bippes, Christian A.; Muller, Daniel J.

    2011-08-01

    Membranes confining cells and cellular compartments are essential for life. Membrane proteins are molecular machines that equip cell membranes with highly sophisticated functionality. Examples of such functions are signaling, ion pumping, energy conversion, molecular transport, specific ligand binding, cell adhesion and protein trafficking. However, it is not well understood how most membrane proteins work and how the living cell regulates their function. We review how atomic force microscopy (AFM) can be applied for structural and functional investigations of native membrane proteins. High-resolution time-lapse AFM imaging records membrane proteins at work, their oligomeric state and their dynamic assembly. The AFM stylus resembles a multifunctional toolbox that allows the measurement of several chemical and physical parameters at the nanoscale. In the single-molecule force spectroscopy (SMFS) mode, AFM quantifies and localizes interactions in membrane proteins that stabilize their folding and modulate their functional state. Dynamic SMFS discloses fascinating insights into the free energy landscape of membrane proteins. Single-cell force spectroscopy quantifies the interactions of live cells with their environment to single-receptor resolution. In the future, technological progress in AFM-based approaches will enable us to study the physical nature of biological interactions in more detail and decipher how cells control basic processes.

  1. Understanding the TERS Effect with On-line Tunneling and Force Feedback Using Multiprobe AFM/NSOM with Raman Integration

    NASA Astrophysics Data System (ADS)

    Lewis, Aaron; Dekhter, Rimma; Hamra, Patricia; Bar-David, Yossi; Taha, Hesham

    Tip enhanced Raman scattering (TERS) has evolved in several directions over the past years. The data from this variety of methodologies has now accumulated to the point that there is a reasonable possibility of evolving an understanding of the underlying cause of the resulting effects that could be the origin of the various TERS enhancement processes. The objective of this presentation is to use the results thus far with atomic force microscopy (AFM) probes with noble metal coating, etching, transparent gold nanoparticles with and without a second nanoparticle [Wang and Schultz, ANALYST 138, 3150 (2013)] and tunneling feedback probes [R. Zhang et. al., NATURE 4 9 8, 8 2 (2013)]. We attempt at understanding this complex of results with AFM/NSOM multiprobe techniques. Results indicate that TERS is dominated by complex quantum interactions. This produces a highly confined and broadband plasmon field with all k vectors for effective excitation. Normal force tuning fork feedback with exposed tip probes provides an excellent means to investigate these effects with TERS probes that we have shown can circumvent the vexing problem of jump to contact prevalent in conventional AFM methodology and permit on-line switching between tunneling and AFM feedback modes of operation.

  2. A practical guide to quantify cell adhesion using single-cell force spectroscopy.

    PubMed

    Friedrichs, Jens; Legate, Kyle R; Schubert, Rajib; Bharadwaj, Mitasha; Werner, Carsten; Müller, Daniel J; Benoit, Martin

    2013-04-01

    Quantitative analysis of cellular interactions with the extracellular environment is necessary to gain an understanding of how cells regulate adhesion in the development and maintenance of multicellular organisms, and how changes in cell adhesion contribute to diseases. We provide a practical guide to quantify the adhesive strength of living animal cells to various substrates using atomic force microscopy (AFM)-based single-cell force spectroscopy (SCFS). We describe how to control cell state and attachment to the AFM cantilever, how to functionalize supports for SCFS measurements, how to conduct cell adhesion measurements, and how to analyze and interpret the recorded SCFS data. This guide is intended to assist newcomers in the field to perform AFM-based SCFS measurements. PMID:23396062

  3. A practical guide to quantify cell adhesion using single-cell force spectroscopy.

    PubMed

    Friedrichs, Jens; Legate, Kyle R; Schubert, Rajib; Bharadwaj, Mitasha; Werner, Carsten; Müller, Daniel J; Benoit, Martin

    2013-04-01

    Quantitative analysis of cellular interactions with the extracellular environment is necessary to gain an understanding of how cells regulate adhesion in the development and maintenance of multicellular organisms, and how changes in cell adhesion contribute to diseases. We provide a practical guide to quantify the adhesive strength of living animal cells to various substrates using atomic force microscopy (AFM)-based single-cell force spectroscopy (SCFS). We describe how to control cell state and attachment to the AFM cantilever, how to functionalize supports for SCFS measurements, how to conduct cell adhesion measurements, and how to analyze and interpret the recorded SCFS data. This guide is intended to assist newcomers in the field to perform AFM-based SCFS measurements.

  4. Mapping site-specific endonuclease binding to DNA by direct imaging with atomic force microscopy (AFM)

    NASA Astrophysics Data System (ADS)

    Allison, David P.; Thundat, Thomas G.; Modrich, P.; Isfort, R. J.; Doktycz, Mitchel J.; Kerper, P. S.; Warmack, R. J.

    1995-04-01

    Physical mapping of DNA can be accomplished by direct AFM imaging of site specific proteins bound to DNA molecules. Using Gln-111, a mutant of EcoRI endonuclease with a specific affinity for EcoRI sites 1000 times greater than wild type enzyme but with cleavage rate constants reduced by a factor of 104, we demonstrate site-specific mapping by direct AFM imaging. Images are presented showing specific-site binding of Gln-111 to plasmids having either one (pBS+) or two (pMP32) EcoRI sites. Identification of the Gln-111/DNA complex is greatly enhanced by biotinylation of the complex followed by reaction with streptavidin gold prior to imaging. Image enhancement coupled with improvements in our preparation techniques for imaging large DNA molecules, such as lambda DNA (47 kb), has the potential to contribute to direct AFM restriction mapping of cosmid-sized genomic DNAs.

  5. Evaluating interaction forces between BSA and rabbit anti-BSA in sulphathiazole sodium, tylosin and levofloxacin solution by AFM

    PubMed Central

    2011-01-01

    Protein-protein interactions play crucial roles in numerous biological processes. However, it is still challenging to evaluate the protein-protein interactions, such as antigen and antibody, in the presence of drug molecules in physiological liquid. In this study, the interaction between bovine serum albumin (BSA) and rabbit anti-BSA was investigated using atomic force microscopy (AFM) in the presence of various antimicrobial drugs (sulphathiazole sodium, tylosin and levofloxacin) under physiological condition. The results show that increasing the concentration of tylosin decreased the single-molecule-specific force between BSA and rabbit anti-BSA. As for sulphathiazole sodium, it dramatically decreased the specific force at a certain critical concentration, but increased the nonspecific force as its concentration increasing. In addition, the presence of levofloxacin did not greatly influence either the specific or nonspecific force. Collectively, these results suggest that these three drugs may adopt different mechanisms to affect the interaction force between BSA and rabbit anti-BSA. These findings may enhance our understanding of antigen/antibody binding processes in the presence of drug molecules, and hence indicate that AFM could be helpful in the design and screening of drugs-modulating protein-protein interaction processes. PMID:22053876

  6. Taking Nanomedicine Teaching into Practice with Atomic Force Microscopy and Force Spectroscopy

    ERIC Educational Resources Information Center

    Carvalho, Filomena A.; Freitas, Teresa; Santos, Nuno C.

    2015-01-01

    Atomic force microscopy (AFM) is a useful and powerful tool to study molecular interactions applied to nanomedicine. The aim of the present study was to implement a hands-on atomic AFM course for graduated biosciences and medical students. The course comprises two distinct practical sessions, where students get in touch with the use of an atomic…

  7. The structure and function of cell membranes examined by atomic force microscopy and single-molecule force spectroscopy.

    PubMed

    Shan, Yuping; Wang, Hongda

    2015-06-01

    The cell membrane is one of the most complicated biological complexes, and long-term fierce debates regarding the cell membrane persist because of technical hurdles. With the rapid development of nanotechnology and single-molecule techniques, our understanding of cell membranes has substantially increased. Atomic force microscopy (AFM) has provided several unprecedented advances (e.g., high resolution, three-dimensional and in situ measurements) in the study of cell membranes and has been used to systematically dissect the membrane structure in situ from both sides of membranes; as a result, novel models of cell membranes have recently been proposed. This review summarizes the new progress regarding membrane structure using in situ AFM and single-molecule force spectroscopy (SMFS), which may shed light on the study of the structure and functions of cell membranes.

  8. Unraveling protein-protein interactions in clathrin assemblies via atomic force spectroscopy.

    PubMed

    Jin, Albert J; Lafer, Eileen M; Peng, Jennifer Q; Smith, Paul D; Nossal, Ralph

    2013-03-01

    Atomic force microscopy (AFM), single molecule force spectroscopy (SMFS), and single particle force spectroscopy (SPFS) are used to characterize intermolecular interactions and domain structures of clathrin triskelia and clathrin-coated vesicles (CCVs). The latter are involved in receptor-mediated endocytosis (RME) and other trafficking pathways. Here, we subject individual triskelia, bovine-brain CCVs, and reconstituted clathrin-AP180 coats to AFM-SMFS and AFM-SPFS pulling experiments and apply novel analytics to extract force-extension relations from very large data sets. The spectroscopic fingerprints of these samples differ markedly, providing important new information about the mechanism of CCV uncoating. For individual triskelia, SMFS reveals a series of events associated with heavy chain alpha-helix hairpin unfolding, as well as cooperative unraveling of several hairpin domains. SPFS of clathrin assemblies exposes weaker clathrin-clathrin interactions that are indicative of inter-leg association essential for RME and intracellular trafficking. Clathrin-AP180 coats are energetically easier to unravel than the coats of CCVs, with a non-trivial dependence on force-loading rate.

  9. Impedance Spectroscopic Investigation of Proton Conductivity in Nafion Using Transient Electrochemical Atomic Force Microscopy (AFM)

    PubMed Central

    Hink, Steffen; Wagner, Norbert; Bessler, Wolfgang G.; Roduner, Emil

    2012-01-01

    Spatially resolved impedance spectroscopy of a Nafion polyelectrolyte membrane is performed employing a conductive and Pt-coated tip of an atomic force microscope as a point-like contact and electrode. The experiment is conducted by perturbing the system by a rectangular voltage step and measuring the incurred current, followed by Fourier transformation and plotting the impedance against the frequency in a conventional Bode diagram. To test the potential and limitations of this novel method, we present a feasibility study using an identical hydrogen atmosphere at a well-defined relative humidity on both sides of the membrane. It is demonstrated that good quality impedance spectra are obtained in a frequency range of 0.2–1000 Hz. The extracted polarization curves exhibit a maximum current which cannot be explained by typical diffusion effects. Simulation based on equivalent circuits requires a Nernst element for restricted diffusion in the membrane which suggests that this effect is based on the potential dependence of the electrolyte resistance in the high overpotential region. PMID:24958175

  10. Spatial dependence of polycrystalline FTO’s conductance analyzed by conductive atomic force microscope (C-AFM)

    SciTech Connect

    Peixoto, Alexandre Pessoa; Costa, J. C. da

    2014-05-15

    Fluorine-doped Tin oxide (FTO) is a highly transparent, electrically conductive polycrystalline material frequently used as an electrode in organic solar cells and optical-electronic devices [1–2]. In this work a spatial analysis of the conductive behavior of FTO was carried out by Conductive-mode Atomic Force Microscopy (C-AFM). Rare highly oriented grains sample give us an opportunity to analyze the top portion of polycrystalline FTO and compare with the border one. It is shown that the current flow essentially takes place through the polycrystalline edge at grain boundaries.

  11. PREFACE: NC-AFM 2005: Proceedings of the 8th International Conference on Non-Contact Atomic Force Microscopy

    NASA Astrophysics Data System (ADS)

    Reichling, M.; Mikosch, W.

    2006-04-01

    The 8th International Conference on Non-Contact Atomic Force Microscopy, held in Bad Essen, Germany, from 15 18th August 2005, attracted a record breaking number of participants presenting excellent contributions from a variety of scientific fields. This clearly demonstrated the high level of activity and innovation present in the community of NC-AFM researchers and the continuous growth of the field. The strongest ever participation of companies for a NC-AFM meeting is a sign for the emergence of new markets for the growing NC-AFM community; and the high standard of the products presented at the exhibition, many of them brand-new developments, reflected the unbroken progress in technology. The development of novel technologies and the sophistication of known techniques in research laboratories and their subsequent commercialization is still a major driving force for progress in this area of nanoscience. The conference was a perfect demonstration of how progress in the development of enabling technologies can readily be transcribed into basic research yielding fundamental insight with an impact across disciplines. The NC-AFM 2005 scientific programme was based on five cornerstones, each representing an area of vivid research and scientific progress. Atomic resolution imaging on oxide surfaces, which has long been a vision for the catalysis community, appears to be routine in several laboratories and after a period of demonstrative experiments NC-AFM now makes unique contributions to the understanding of processes in surface chemistry. These capabilities also open up new routes for the analysis of clusters and molecules deposited on dielectric surfaces where resolution limits are pushed towards the single atom level. Atomic precision manipulation with the dynamic AFM left the cradle of its infancy and flourishes in the family of bottom-up fabrication nanotechnologies. The systematic development of established and the introduction of new concepts of contrast

  12. Interaction and dynamics of ambient water adlayers on graphite probed using AFM voltage nanolithography and electrostatic force microscopy.

    PubMed

    Gowthami, T; Kurra, Narendra; Raina, Gargi

    2014-04-18

    In this work, we report the impact of the interaction and dynamics of increasing ambient water adlayers on etch patterns on a hydrophobic highly oriented pyrolytic graphite (HOPG) surface obtained using atomic force microscopy (AFM) voltage nanolithography in contact mode by applying a positive bias to the sample. The changes in the dimensions of the etch patterns were investigated as a function of the increasing number of water adlayers present on the HOPG, which is varied by changing the time interval since HOPG cleavage. Changes in the width of the etch patterns and the surrounding water droplets were monitored with time, using intermittent-contact-mode AFM. Electrostatic force microscopy (EFM) has been employed to study the charged nature of the etch patterns and the neighboring water film with time. The width of the etch patterns made on freshly cleaved HOPG shows an increase of ∼33% over 48 h, whereas nine-day-old cleaved HOPG shows a 79% increase over the same period. No changes in the dimensions are observed while imaging in a nitrogen atmosphere soon after lithography. In ambient conditions, the EFM phase shift of the patterns shows a large change of ∼84-88% over 30 h. This study demonstrates the effect of the stored electrostatic energy of a polarized ice-like water adlayer, resulting in changes in the dimensions of the etch patterns long after lithography, whereas liquid-like water droplets do not affect the etch patterns.

  13. Interaction and dynamics of ambient water adlayers on graphite probed using AFM voltage nanolithography and electrostatic force microscopy.

    PubMed

    Gowthami, T; Kurra, Narendra; Raina, Gargi

    2014-04-18

    In this work, we report the impact of the interaction and dynamics of increasing ambient water adlayers on etch patterns on a hydrophobic highly oriented pyrolytic graphite (HOPG) surface obtained using atomic force microscopy (AFM) voltage nanolithography in contact mode by applying a positive bias to the sample. The changes in the dimensions of the etch patterns were investigated as a function of the increasing number of water adlayers present on the HOPG, which is varied by changing the time interval since HOPG cleavage. Changes in the width of the etch patterns and the surrounding water droplets were monitored with time, using intermittent-contact-mode AFM. Electrostatic force microscopy (EFM) has been employed to study the charged nature of the etch patterns and the neighboring water film with time. The width of the etch patterns made on freshly cleaved HOPG shows an increase of ∼33% over 48 h, whereas nine-day-old cleaved HOPG shows a 79% increase over the same period. No changes in the dimensions are observed while imaging in a nitrogen atmosphere soon after lithography. In ambient conditions, the EFM phase shift of the patterns shows a large change of ∼84-88% over 30 h. This study demonstrates the effect of the stored electrostatic energy of a polarized ice-like water adlayer, resulting in changes in the dimensions of the etch patterns long after lithography, whereas liquid-like water droplets do not affect the etch patterns. PMID:24651210

  14. Force profiles of protein pulling with or without cytoskeletal links studied by AFM

    SciTech Connect

    Afrin, Rehana; Ikai, Atsushi . E-mail: aikai@bio.titech.ac.jp

    2006-09-15

    To test the capability of the atomic force microscope for distinguishing membrane proteins with/without cytoskeletal associations, we studied the pull-out mechanics of lipid tethers from the red blood cell (RBC). When wheat germ agglutinin, a glycophorin A (GLA) specific lectin, was used to pull out tethers from RBC, characteristic force curves for tether elongation having a long plateau force were observed but without force peaks which are usually attributed to the forced unbinding of membrane components from the cytoskeleton. The result was in agreement with the reports that GLA is substantially free of cytoskeletal interactions. On the contrary, when the Band 3 specific lectin, concanavalin A, was used, the force peaks were indeed observed together with a plateau supporting its reported cytoskeletal association. Based on these observations, we postulate that the state of cytoskeletal association of particular membrane proteins can be identified from the force profiles of their pull-out mechanics.

  15. Direct and quantitative AFM measurements of the concentration and temperature dependence of the hydrophobic force law at nanoscopic contacts.

    PubMed

    Stock, Philipp; Utzig, Thomas; Valtiner, Markus

    2015-05-15

    By virtue of its importance for self-organization of biological matter the hydrophobic force law and the range of hydrophobic interactions (HI) have been debated extensively over the last 40 years. Here, we directly measure and quantify the hydrophobic force-distance law over large temperature and concentration ranges. In particular, we study the HI between molecularly smooth hydrophobic self-assembled monolayers, and similarly modified gold-coated AFM tips (radii∼8-50 nm). We present quantitative and direct evidence that the hydrophobic force is both long-ranged and exponential down to distances of about 1-2 nm. Therefore, we introduce a self-consistent radius-normalization for atomic force microscopy data. This approach allows quantitative data fitting of AFM-based experimental data to the recently proposed Hydra-model. With a statistical significance of r(2)⩾0.96 our fitting and data directly reveal an exponential HI decay length of 7.2±1.2 Å that is independent of the salt concentration up to 750 mM. As such, electrostatic screening does not have a significant influence on the HI in electrolyte concentrations ranging from 1 mM to 750 mM. In 1 M solutions the observed instability during approach shifts to longer distances, indicating ion correlation/adsorption effects at high salt concentrations. With increasing temperature the magnitude of HI decreases monotonically, while the range increases slightly. We compare our results to the large body of available literature, and shed new light into range and magnitude of hydrophobic interactions at very close distances and over wide temperature and concentration regimes. PMID:25678158

  16. Self assembly of epicuticular waxes on living plant surfaces imaged by atomic force microscopy (AFM).

    PubMed

    Koch, Kerstin; Neinhuis, Christoph; Ensikat, Hans-Jürgen; Barthlott, Wilhelm

    2004-03-01

    The cuticle of terrestrial vascular plants and some bryophytes is covered with a complex mixture of lipids, usually called epicuticular waxes. Self-assembly processes of wax molecules lead to crystalline three-dimensional micro- and nanostructures that emerge from an underlying wax film. This paper presents the first AFM study on wax regeneration on the surfaces of living plants and the very early stages of wax crystal formation at the molecular level. Wax formation was analysed on the leaves of Euphorbia lathyris, Galanthus nivalis, and Ipheion uniflorum. Immediately after wax removal, regeneration of a wax film began, consisting of individual layers of, typically, 3-5 nm thickness. Subsequently, several different stages of crystal growth could be distinguished, and different patterns of wax regeneration as well as considerable variation in regeneration speed were found.

  17. PREFACE: NC-AFM 2006: Proceedings of the 9th International Conference on Non-contact Atomic Force Microscopy

    NASA Astrophysics Data System (ADS)

    Tomitori, Masahiko; Onishi, Hiroshi

    2007-02-01

    The advent of scanning probe microscopy (SPM) in the 1980s has significantly promoted nanoscience and nanotechnology. In particular, non-contact atomic force microscopy (NC-AFM), one of the SPM family, has unique capabilities with high spatial resolution for nanoscale measurements in vacuum, air and liquids. In the last decade we have witnessed the rapid progress of NC-AFM with improved performance and increasing applications. A series of NC-AFM international conferences have greatly contributed to this field. Initiated in Osaka in 1998, the NC-AFM meeting has been followed by annual conferences at Pontresina, Hamburg, Kyoto, Montreal, Dingle, Seattle and Bad Essen. The 9th conference was held in Kobe, Japan, 16-20 July 2006. This special issue of Nanotechnology contains the outstanding contributions of the conference. During the meeting delegates learnt about a number of significant advances. Topics covered atomic resolution imaging of metals, semiconductors, insulators, ionic crystals, oxides, molecular systems, imaging of biological materials in various environments and novel instrumentation. Work also included the characterization of electronic and magnetic properties, tip and cantilever fabrication and characterization, atomic distinction based on analysis of tip-sample interaction, atomic scale manipulation, fabrication of nanostructures using NC-AFM, and related theories and simulations. We are greatly impressed by the increasing number of applications, and convinced that NC-AFM and related techniques are building a bridge to a future nano world, where quantum phenomena will dominate and nano devices will be realized. In addition, a special session on SPM road maps was held as a first trial in the field, where the future prospects of SPM were discussed enthusiastically. The overall success of the NC-AFM 2006 conference was due to the efforts of many individuals and groups with respect to scientific and technological progress, as well as the international

  18. Recording the dynamic endocytosis of single gold nanoparticles by AFM-based force tracing.

    PubMed

    Ding, Bohua; Tian, Yongmei; Pan, Yangang; Shan, Yuping; Cai, Mingjun; Xu, Haijiao; Sun, Yingchun; Wang, Hongda

    2015-05-01

    We utilized force tracing to directly record the endocytosis of single gold nanoparticles (Au NPs) with different sizes, revealing the size-dependent endocytosis dynamics and the crucial role of membrane cholesterol. The force, duration and velocity of Au NP invagination are accurately determined at the single-particle and microsecond level unprecedentedly. PMID:25864702

  19. Nanopipette combined with quartz tuning fork-atomic force microscope for force spectroscopy/microscopy and liquid delivery-based nanofabrication

    SciTech Connect

    An, Sangmin; Lee, Kunyoung; Kim, Bongsu; Noh, Haneol; Kim, Jongwoo; Kwon, Soyoung; Lee, Manhee; Hong, Mun-Heon; Jhe, Wonho

    2014-03-15

    This paper introduces a nanopipette combined with a quartz tuning fork-atomic force microscope system (nanopipette/QTF-AFM), and describes experimental and theoretical investigations of the nanoscale materials used. The system offers several advantages over conventional cantilever-based AFM and QTF-AFM systems, including simple control of the quality factor based on the contact position of the QTF, easy variation of the effective tip diameter, electrical detection, on-demand delivery and patterning of various solutions, and in situ surface characterization after patterning. This tool enables nanoscale liquid delivery and nanofabrication processes without damaging the apex of the tip in various environments, and also offers force spectroscopy and microscopy capabilities.

  20. Nanopipette combined with quartz tuning fork-atomic force microscope for force spectroscopy/microscopy and liquid delivery-based nanofabrication.

    PubMed

    An, Sangmin; Lee, Kunyoung; Kim, Bongsu; Noh, Haneol; Kim, Jongwoo; Kwon, Soyoung; Lee, Manhee; Hong, Mun-Heon; Jhe, Wonho

    2014-03-01

    This paper introduces a nanopipette combined with a quartz tuning fork-atomic force microscope system (nanopipette/QTF-AFM), and describes experimental and theoretical investigations of the nanoscale materials used. The system offers several advantages over conventional cantilever-based AFM and QTF-AFM systems, including simple control of the quality factor based on the contact position of the QTF, easy variation of the effective tip diameter, electrical detection, on-demand delivery and patterning of various solutions, and in situ surface characterization after patterning. This tool enables nanoscale liquid delivery and nanofabrication processes without damaging the apex of the tip in various environments, and also offers force spectroscopy and microscopy capabilities.

  1. Recording the dynamic endocytosis of single gold nanoparticles by AFM-based force tracing

    NASA Astrophysics Data System (ADS)

    Ding, Bohua; Tian, Yongmei; Pan, Yangang; Shan, Yuping; Cai, Mingjun; Xu, Haijiao; Sun, Yingchun; Wang, Hongda

    2015-04-01

    We utilized force tracing to directly record the endocytosis of single gold nanoparticles (Au NPs) with different sizes, revealing the size-dependent endocytosis dynamics and the crucial role of membrane cholesterol. The force, duration and velocity of Au NP invagination are accurately determined at the single-particle and microsecond level unprecedentedly.We utilized force tracing to directly record the endocytosis of single gold nanoparticles (Au NPs) with different sizes, revealing the size-dependent endocytosis dynamics and the crucial role of membrane cholesterol. The force, duration and velocity of Au NP invagination are accurately determined at the single-particle and microsecond level unprecedentedly. Electronic supplementary information (ESI) available: Details of the experimental procedures and the results of the control experiments. See DOI: 10.1039/c5nr01020a

  2. Development and testing of hyperbaric atomic force microscopy (AFM) and fluorescence microscopy for biological applications.

    PubMed

    D'Agostino, D P; McNally, H A; Dean, J B

    2012-05-01

    A commercially available atomic force microscopy and fluorescence microscope were installed and tested inside a custom-designed hyperbaric chamber to provide the capability to study the effects of hyperbaric gases on biological preparations, including cellular mechanism of oxidative stress. In this report, we list details of installing and testing atomic force microscopy and fluorescence microscopy inside a hyperbaric chamber. The pressure vessel was designed to accommodate a variety of imaging equipment and ensures full functionality at ambient and hyperbaric conditions (≤85 psi). Electrical, gas and fluid lines were installed to enable remote operation of instrumentation under hyperbaric conditions, and to maintain viable biological samples with gas-equilibrated superfusate and/or drugs. Systems were installed for vibration isolation and temperature regulation to maintain atomic force microscopy performance during compression and decompression. Results of atomic force microscopy testing demonstrate sub-nanometre resolution at hyperbaric pressure in dry scans and fluid scans, in both contact mode and tapping mode. Noise levels were less when measurements were taken under hyperbaric pressure with air, helium (He) and nitrogen (N(2) ). Atomic force microscopy and fluorescence microscopy measurements were made on a variety of living cell cultures exposed to hyperbaric gases (He, N(2) , O(2) , air). In summary, atomic force microscopy and fluorescence microscopy were installed and tested for use at hyperbaric pressures and enables the study of cellular and molecular effects of hyperbaric gases and pressure per se in biological preparations.

  3. Single Cell Wall Nonlinear Mechanics Revealed by a Multiscale Analysis of AFM Force-Indentation Curves.

    PubMed

    Digiuni, Simona; Berne-Dedieu, Annik; Martinez-Torres, Cristina; Szecsi, Judit; Bendahmane, Mohammed; Arneodo, Alain; Argoul, Françoise

    2015-05-01

    Individual plant cells are rather complex mechanical objects. Despite the fact that their wall mechanical strength may be weakened by comparison with their original tissue template, they nevertheless retain some generic properties of the mother tissue, namely the viscoelasticity and the shape of their walls, which are driven by their internal hydrostatic turgor pressure. This viscoelastic behavior, which affects the power-law response of these cells when indented by an atomic force cantilever with a pyramidal tip, is also very sensitive to the culture media. To our knowledge, we develop here an original analyzing method, based on a multiscale decomposition of force-indentation curves, that reveals and quantifies for the first time the nonlinearity of the mechanical response of living single plant cells upon mechanical deformation. Further comparing the nonlinear strain responses of these isolated cells in three different media, we reveal an alteration of their linear bending elastic regime in both hyper- and hypotonic conditions. PMID:25954881

  4. Single Cell Wall Nonlinear Mechanics Revealed by a Multiscale Analysis of AFM Force-Indentation Curves

    PubMed Central

    Digiuni, Simona; Berne-Dedieu, Annik; Martinez-Torres, Cristina; Szecsi, Judit; Bendahmane, Mohammed; Arneodo, Alain; Argoul, Françoise

    2015-01-01

    Individual plant cells are rather complex mechanical objects. Despite the fact that their wall mechanical strength may be weakened by comparison with their original tissue template, they nevertheless retain some generic properties of the mother tissue, namely the viscoelasticity and the shape of their walls, which are driven by their internal hydrostatic turgor pressure. This viscoelastic behavior, which affects the power-law response of these cells when indented by an atomic force cantilever with a pyramidal tip, is also very sensitive to the culture media. To our knowledge, we develop here an original analyzing method, based on a multiscale decomposition of force-indentation curves, that reveals and quantifies for the first time the nonlinearity of the mechanical response of living single plant cells upon mechanical deformation. Further comparing the nonlinear strain responses of these isolated cells in three different media, we reveal an alteration of their linear bending elastic regime in both hyper- and hypotonic conditions. PMID:25954881

  5. Dielectrophoresis force spectroscopy for colloidal clusters.

    PubMed

    Park, Hyunjoo; Wei, Ming-Tzo; Ou-Yang, H Daniel

    2012-08-01

    Optical trapping-based force spectroscopy was used to measure the frequency-dependent DEP forces and DEP crossover frequencies of colloidal polymethyl methacrylate spheres and clusters. A single sphere or cluster, held by an optical tweezer, was positioned near the center of a pair of gold-film electrodes where alternating current elecroosmosis flow was negligible. Use of amplitude modulation and phase-sensitive lock-in detection for accurate measurement of the DEP force yielded new insight into dielectric relaxation mechanisms near the crossover frequencies. On one hand, the size dependence of the DEP force near the crossover frequencies indicates that the dominant polarization mechanism is a volume effect. On the other hand, the power-law dependence of the crossover frequency on the particle radius with an exponent of -2 indicates the dielectric relaxation is more likely because of ionic diffusion across the particle surface, suggesting the dominant polarization mechanism may be a surface polarization effect. Better theories are needed to explain the experiment. Nevertheless, the strong size dependence of the crossover frequencies suggests the use of DEP for size sorting of micron-sized particles. PMID:22899256

  6. Direct AFM force measurements between air bubbles in aqueous monodisperse sodium poly(styrene sulfonate) solutions.

    PubMed

    Browne, Christine; Tabor, Rico F; Grieser, Franz; Dagastine, Raymond R

    2015-08-01

    Structural forces play an important role in the rheology, processing and stability of colloidal systems and complex fluids, with polyelectrolytes representing a key class of structuring colloids. Here, we explore the interactions between soft colloids, in the form of air bubbles, in solutions of monodisperse sodium poly(styrene sulfonate) as a model polyelectrolyte. It is found that by self-consistently modelling the force oscillations due to structuring of the polymer chains along with deformation of the bubbles, it is possible to precisely predict the interaction potential between approaching bubbles. In line with polyelectrolyte scaling theory, two distinct regimes of behaviour are seen, corresponding to dilute and semi-dilute polymer solutions. It is also seen that by blending monodisperse systems to give a bidisperse sample, the interaction forces between soft colloids can be controlled with a high degree of precision. At increasing bubble collision velocity, it is revealed that hydrodynamic flow overwhelms oscillatory structural interactions, showing the important disparity between equilibrium behaviour and dynamic interactions.

  7. Double Layer of a Gold Electrode Probed by AFM Force Measurements.

    PubMed

    Barten, D; Kleijn, J M; Duval, J; Leeuwen, H P V; Lyklema, J; Cohen Stuart, M A

    2003-02-18

    Colloidal probe atomic force microscopy was used to determine the electric double layer interactions between a gold electrode and a spherical silica probe. The double layer properties of the gold/solution interface were varied through the pH and salt concentration of the electrolyte, as well as by externally applying an electric potential. The double layer potentials ψ(d) of the gold surface were obtained by fitting the force-distance curves according to the DLVO (Derjaguin-Landau-Verwey-Overbeek) theory, using earlier obtained values for the double layer potential of the silica probe as input parameter. It was found that the gold electrode combines the features of reversible and polarizable interfaces; i.e., its charge and potential are determined by both the solution pH and the external potential. The pH dependence is attributed to proton adsorption and desorption from oxidic groups on the gold surface. In the potential range studied, ψ(d) varies linearly with the applied potential; the variation in ψ(d) is roughly 10% of that in the applied potential. The potential of zero force (the external potential at which ψ(d) = 0) varies with pH. The various features of the gold/electrolyte interface are described well by an amphifunctional double layer model. The results of this study form the basis of the interpretation of adsorption studies on gold as a function of pH and externally applied potential.

  8. An integrated methodology for data processing in dynamic force spectroscopy of ligand-receptor binding.

    PubMed

    Odorico, M; Teulon, J-M; Berthoumieu, O; Chen, S-w W; Parot, P; Pellequer, J-L

    2007-10-01

    Dynamic force spectroscopy (DFS), using atomic force microscopy (AFM), is a powerful tool to study ligand-receptor binding. The interaction mode of two binding partners is investigated by exploring stochastic behaviors of bond rupture events. However, to define a rupture event from force-distance measurements is not conclusive or unique in literature. To reveal the influence of event identification methods, we have developed an efficient protocol to manage tremendous amount of data by implementing different choices of peak selection from the force-distance curve. This data processing software simplifies routinely experimental procedures such as cantilever spring constant and force-distance curve calibrations, statistical treatments of data, and analysis distributions of rupture events. In the present work, we took available experimental data from a complex between a chelate metal compound and a monoclonal antibody as a study system.

  9. Stimulated Raman spectroscopy and nanoscopy of molecules using near field photon induced forces without resonant electronic enhancement gain

    NASA Astrophysics Data System (ADS)

    Tamma, Venkata Ananth; Huang, Fei; Nowak, Derek; Kumar Wickramasinghe, H.

    2016-06-01

    We report on stimulated Raman spectroscopy and nanoscopy of molecules, excited without resonant electronic enhancement gain, and recorded using near field photon induced forces. Photon-induced interaction forces between the sharp metal coated silicon tip of an Atomic Force Microscope (AFM) and a sample resulting from stimulated Raman excitation were detected. We controlled the tip to sample spacing using the higher order flexural eigenmodes of the AFM cantilever, enabling the tip to come very close to the sample. As a result, the detection sensitivity was increased compared with previous work on Raman force microscopy. Raman vibrational spectra of azobenzene thiol and l-phenylalanine were measured and found to agree well with published results. Near-field force detection eliminates the need for far-field optical spectrometer detection. Recorded images show spatial resolution far below the optical diffraction limit. Further optimization and use of ultrafast pulsed lasers could push the detection sensitivity towards the single molecule limit.

  10. Attractive forces between hydrophobic solid surfaces measured by AFM on the first approach in salt solutions and in the presence of dissolved gases.

    PubMed

    Azadi, Mehdi; Nguyen, Anh V; Yakubov, Gleb E

    2015-02-17

    Interfacial gas enrichment of dissolved gases (IGE) has been shown to cover hydrophobic solid surfaces in water. The atomic force microscopy (AFM) data has recently been supported by molecular dynamics simulation. It was demonstrated that IGE is responsible for the unexpected stability and large contact angle of gaseous nanobubbles at the hydrophobic solid-water interface. Here we provide further evidence of the significant effect of IGE on an attractive force between hydrophobic solid surfaces in water. The force in the presence of dissolved gas, i.e., in aerated and nonaerated NaCl solutions (up to 4 M), was measured by the AFM colloidal probe technique. The effect of nanobubble bridging on the attractive force was minimized or eliminated by measuring forces on the first approach of the AFM probe toward the flat hydrophobic surface and by using high salt concentrations to reduce gas solubility. Our results confirm the presence of three types of forces, two of which are long-range attractive forces of capillary bridging origin as caused by either surface nanobubbles or gap-induced cavitation. The third type is a short-range attractive force observed in the absence of interfacial nanobubbles that is attributed to the IGE in the form of a dense gas layer (DGL) at hydrophobic surfaces. Such a force was found to increase with increasing gas saturation and to decrease with decreasing gas solubility.

  11. Dielectric spectroscopy at the nanoscale by atomic force microscopy: A simple model linking materials properties and experimental response

    SciTech Connect

    Miccio, Luis A. Colmenero, Juan; Kummali, Mohammed M.; Alegría, Ángel; Schwartz, Gustavo A.

    2014-05-14

    The use of an atomic force microscope for studying molecular dynamics through dielectric spectroscopy with spatial resolution in the nanometer scale is a recently developed approach. However, difficulties in the quantitative connection of the obtained data and the material dielectric properties, namely, frequency dependent dielectric permittivity, have limited its application. In this work, we develop a simple electrical model based on physically meaningful parameters to connect the atomic force microscopy (AFM) based dielectric spectroscopy experimental results with the material dielectric properties. We have tested the accuracy of the model and analyzed the relevance of the forces arising from the electrical interaction with the AFM probe cantilever. In this way, by using this model, it is now possible to obtain quantitative information of the local dielectric material properties in a broad frequency range. Furthermore, it is also possible to determine the experimental setup providing the best sensitivity in the detected signal.

  12. Characterization of gold nanoparticle films: Rutherford backscattering spectroscopy, scanning electron microscopy with image analysis, and atomic force microscopy

    SciTech Connect

    Lansåker, Pia C. Niklasson, Gunnar A.; Granqvist, Claes G.; Hallén, Anders

    2014-10-15

    Gold nanoparticle films are of interest in several branches of science and technology, and accurate sample characterization is needed but technically demanding. We prepared such films by DC magnetron sputtering and recorded their mass thickness by Rutherford backscattering spectroscopy. The geometric thickness d{sub g}—from the substrate to the tops of the nanoparticles—was obtained by scanning electron microscopy (SEM) combined with image analysis as well as by atomic force microscopy (AFM). The various techniques yielded an internally consistent characterization of the films. In particular, very similar results for d{sub g} were obtained by SEM with image analysis and by AFM.

  13. AFM in peak force mode applied to worn siloxane-hydrogel contact lenses.

    PubMed

    Abadías, Clara; Serés, Carme; Torrent-Burgués, Juan

    2015-04-01

    The objective of this work is to apply Atomic Force Microscopy in Peak Force mode to obtain topographic characteristics (mean roughness, root-mean-square roughness, skewness and kurtosis) and mechanical characteristics (adhesion, elastic modulus) of Siloxane-Hydrogel Soft Contact Lenses (CLs) of two different materials, Lotrafilcon B of Air Optix (AO) and Asmofilcon A of PremiO (P), after use (worn CLs). Thus, the results obtained with both materials will be compared, as well as the changes produced by the wear at a nanoscopic level. The results show significant changes in the topographic and mechanical characteristics of the CLs, at a nanoscopic level, due to wear. The AO CL show values of the topographic parameters lower than those of the P CL after wear, which correlates with a better comfort qualification given to the former by the wearers. A significant correlation has also been obtained between the adhesion values found after the use of the CLs with tear quality tests, both break-up-time and Schirmer.

  14. High-resolution noncontact AFM and Kelvin probe force microscopy investigations of self-assembled photovoltaic donor–acceptor dyads

    PubMed Central

    Schwartz, Pierre-Olivier; Biniek, Laure; Brinkmann, Martin; Leclerc, Nicolas; Zaborova, Elena

    2016-01-01

    Summary Self-assembled donor–acceptor dyads are used as model nanostructured heterojunctions for local investigations by noncontact atomic force microscopy (nc-AFM) and Kelvin probe force microscopy (KPFM). With the aim to probe the photo-induced charge carrier generation, thin films deposited on transparent indium tin oxide substrates are investigated in dark conditions and upon illumination. The topographic and contact potential difference (CPD) images taken under dark conditions are analysed in view of the results of complementary transmission electron microscopy (TEM) experiments. After in situ annealing, it is shown that the dyads with longer donor blocks essentially lead to standing acceptor–donor lamellae, where the acceptor and donor groups are π-stacked in an edge-on configuration. The existence of strong CPD and surface photo-voltage (SPV) contrasts shows that structural variations occur within the bulk of the edge-on stacks. SPV images with a very high lateral resolution are achieved, which allows for the resolution of local photo-charging contrasts at the scale of single edge-on lamella. This work paves the way for local investigations of the optoelectronic properties of donor–acceptor supramolecular architectures down to the elementary building block level. PMID:27335768

  15. High-resolution noncontact AFM and Kelvin probe force microscopy investigations of self-assembled photovoltaic donor-acceptor dyads.

    PubMed

    Grévin, Benjamin; Schwartz, Pierre-Olivier; Biniek, Laure; Brinkmann, Martin; Leclerc, Nicolas; Zaborova, Elena; Méry, Stéphane

    2016-01-01

    Self-assembled donor-acceptor dyads are used as model nanostructured heterojunctions for local investigations by noncontact atomic force microscopy (nc-AFM) and Kelvin probe force microscopy (KPFM). With the aim to probe the photo-induced charge carrier generation, thin films deposited on transparent indium tin oxide substrates are investigated in dark conditions and upon illumination. The topographic and contact potential difference (CPD) images taken under dark conditions are analysed in view of the results of complementary transmission electron microscopy (TEM) experiments. After in situ annealing, it is shown that the dyads with longer donor blocks essentially lead to standing acceptor-donor lamellae, where the acceptor and donor groups are π-stacked in an edge-on configuration. The existence of strong CPD and surface photo-voltage (SPV) contrasts shows that structural variations occur within the bulk of the edge-on stacks. SPV images with a very high lateral resolution are achieved, which allows for the resolution of local photo-charging contrasts at the scale of single edge-on lamella. This work paves the way for local investigations of the optoelectronic properties of donor-acceptor supramolecular architectures down to the elementary building block level. PMID:27335768

  16. Mechanical design of proteins studied by single-molecule force spectroscopy and protein engineering.

    PubMed

    Carrion-Vazquez, M; Oberhauser, A F; Fisher, T E; Marszalek, P E; Li, H; Fernandez, J M

    2000-01-01

    Mechanical unfolding and refolding may regulate the molecular elasticity of modular proteins with mechanical functions. The development of the atomic force microscopy (AFM) has recently enabled the dynamic measurement of these processes at the single-molecule level. Protein engineering techniques allow the construction of homomeric polyproteins for the precise analysis of the mechanical unfolding of single domains. alpha-Helical domains are mechanically compliant, whereas beta-sandwich domains, particularly those that resist unfolding with backbone hydrogen bonds between strands perpendicular to the applied force, are more stable and appear frequently in proteins subject to mechanical forces. The mechanical stability of a domain seems to be determined by its hydrogen bonding pattern and is correlated with its kinetic stability rather than its thermodynamic stability. Force spectroscopy using AFM promises to elucidate the dynamic mechanical properties of a wide variety of proteins at the single molecule level and provide an important complement to other structural and dynamic techniques (e.g., X-ray crystallography, NMR spectroscopy, patch-clamp). PMID:11106807

  17. Combined low-temperature scanning tunneling/atomic force microscope for atomic resolution imaging and site-specific force spectroscopy

    SciTech Connect

    Schwarz, Udo; Albers, Boris J.; Liebmann, Marcus; Schwendemann, Todd C.; Baykara, Mehmet Z.; Heyde, Markus; Salmeron, Miquel; Altman, Eric I.; Schwarz, Udo D.

    2008-02-27

    The authors present the design and first results of a low-temperature, ultrahigh vacuum scanning probe microscope enabling atomic resolution imaging in both scanning tunneling microscopy (STM) and noncontact atomic force microscopy (NC-AFM) modes. A tuning-fork-based sensor provides flexibility in selecting probe tip materials, which can be either metallic or nonmetallic. When choosing a conducting tip and sample, simultaneous STM/NC-AFM data acquisition is possible. Noticeable characteristics that distinguish this setup from similar systems providing simultaneous STM/NC-AFM capabilities are its combination of relative compactness (on-top bath cryostat needs no pit), in situ exchange of tip and sample at low temperatures, short turnaround times, modest helium consumption, and unrestricted access from dedicated flanges. The latter permits not only the optical surveillance of the tip during approach but also the direct deposition of molecules or atoms on either tip or sample while they remain cold. Atomic corrugations as low as 1 pm could successfully be resolved. In addition, lateral drifts rates of below 15 pm/h allow long-term data acquisition series and the recording of site-specific spectroscopy maps. Results obtained on Cu(111) and graphite illustrate the microscope's performance.

  18. Iterative control approach to high-speed force-distance curve measurement using AFM: time-dependent response of PDMS example.

    PubMed

    Kim, Kyong-Soo; Lin, Zhiqun; Shrotriya, Pranav; Sundararajan, Sriram; Zou, Qingze

    2008-08-01

    Force-distance curve measurements using atomic force microscope (AFM) has been widely used in a broad range of areas. However, currently force-curve measurements are hampered the its low speed of AFM. In this article, a novel inversion-based iterative control technique is proposed to dramatically increase the speed of force-curve measurements. Experimental results are presented to show that by using the proposed control technique, the speed of force-curve measurements can be increased by over 80 times--with no loss of spatial resolution--on a commercial AFM platform and with a standard cantilever. High-speed force curve measurements using this control technique are utilized to quantitatively study the time-dependent elastic modulus of poly(dimethylsiloxane) (PDMS). The force-curves employ a broad spectrum of push-in (load) rates, spanning two-order differences. The elastic modulus measured at low-speed compares well with the value obtained from dynamic mechanical analysis (DMA) test, and the value of the elastic modulus increases as the push-in rate increases, signifying that a faster external deformation rate transitions the viscoelastic response of PDMS from that of a rubbery material toward a glassy one. PMID:18467033

  19. Hands-on Force Spectroscopy: Weird Springs and Protein Folding

    ERIC Educational Resources Information Center

    Euler, Manfred

    2008-01-01

    A force spectroscopy model experiment is presented using a low-cost tensile apparatus described earlier. Force-extension measurements of twisted rubber bands are obtained. They exhibit a complex nonlinear elastic behaviour that resembles atomic force spectroscopy investigations of molecules of titin, a muscle protein. The model experiments open up…

  20. Sample stage designed for force modulation microscopy using a tip-mounted AFM scanner.

    PubMed

    Lu, Lu; Xu, Song; Zhang, Donghui; Garno, Jayne C

    2016-03-01

    Among the modes of scanning probe microscopy (SPM), force modulation microscopy (FMM) is often used to acquire mechanical properties of samples concurrent with topographic information. The FMM mode is useful for investigations with polymer and organic thin film samples. Qualitative evaluation of the mixed domains of co-polymers or composite films can often be accomplished with high resolution using FMM phase and amplitude images. We have designed and tested a sample stage for FMM constructed of machined polycarbonate. A generic design enables FMM experiments for instrument configurations with a tip-mounted SPM scanner. A piezoactuator within the sample stage was used to drive the sample to vibrate in the z-direction according to selected parameters. To evaluate the FMM sample stage, we tested samples of known composition with nanoscale dimensions for increasingly complex surface morphologies. Excellent resolution was achieved in ambient conditions using the home-constructed sample stage, as revealed for complex surfaces or multi-component samples. Test structures of nanoholes within a film of organosilanes provided the simplest platform with two distinct surface domains. Ring-shaped nanostructures prepared on Si(111) with mixed organosilanes provided three regions for evaluating FMM results. A complex sample consisting of a cyclic gel polymer containing fibril nanostructures was also tested with FMM measurements. Frequency spectra were acquired for sample domains, revealing distinct differences in local mechanical response. We demonstrate a practical approach to construct a sample stage accessory to facilitate z-sample modulation for FMM experiments with tip-mounted SPM scanners. PMID:26824091

  1. Dissipation and oscillatory solvation forces in confined liquids studied by small-amplitude atomic force spectroscopy.

    PubMed

    de Beer, Sissi; van den Ende, Dirk; Mugele, Frieder

    2010-08-13

    We determine conservative and dissipative tip-sample interaction forces from the amplitude and phase response of acoustically driven atomic force microscope (AFM) cantilevers using a non-polar model fluid (octamethylcyclotetrasiloxane, which displays strong molecular layering) and atomically flat surfaces of highly ordered pyrolytic graphite. Taking into account the base motion and the frequency-dependent added mass and hydrodynamic damping on the AFM cantilever, we develop a reliable force inversion procedure that allows for extracting tip-sample interaction forces for a wide range of drive frequencies. We systematically eliminate the effect of finite drive amplitudes. Dissipative tip-sample forces are consistent with the bulk viscosity down to a thickness of 2-3 nm. Dissipation measurements far below resonance, which we argue to be the most reliable, indicate the presence of peaks in the damping, corresponding to an enhanced 'effective' viscosity, upon expelling the last and second-last molecular layer. PMID:20639584

  2. Single-molecule force spectroscopy: optical tweezers, magnetic tweezers and atomic force microscopy

    PubMed Central

    Neuman, Keir C.; Nagy, Attila

    2012-01-01

    Single-molecule force spectroscopy has emerged as a powerful tool to investigate the forces and motions associated with biological molecules and enzymatic activity. The most common force spectroscopy techniques are optical tweezers, magnetic tweezers and atomic force microscopy. These techniques are described and illustrated with examples highlighting current capabilities and limitations. PMID:18511917

  3. AFM force measurements of the gp120-sCD4 and gp120 or CD4 antigen-antibody interactions

    SciTech Connect

    Chen, Yong; Zeng, Gucheng; Chen, Sherry Shiyi; Feng, Qian; Chen, Zheng Wei

    2011-04-08

    Highlights: {yields} The unbinding force of sCD4-gp120 interaction was 25.45 {+-} 20.46 pN. {yields} The unbinding force of CD4 antigen-antibody interaction was 51.22 {+-} 34.64 pN. {yields} The unbinding force of gp120 antigen-antibody interaction was 89.87 {+-} 44.63 pN. {yields} The interaction forces between various HIV inhibitors and the target molecules are significantly different. {yields} Functionalizing on AFM tip or substrate of an interaction pair caused different results. -- Abstract: Soluble CD4 (sCD4), anti-CD4 antibody, and anti-gp120 antibody have long been regarded as entry inhibitors in human immunodeficiency virus (HIV) therapy. However, the interactions between these HIV entry inhibitors and corresponding target molecules are still poorly understood. In this study, atomic force microscopy (AFM) was utilized to investigate the interaction forces among them. We found that the unbinding forces of sCD4-gp120 interaction, CD4 antigen-antibody interaction, and gp120 antigen-antibody interaction were 25.45 {+-} 20.46, 51.22 {+-} 34.64, and 89.87 {+-} 44.63 pN, respectively, which may provide important mechanical information for understanding the effects of viral entry inhibitors on HIV infection. Moreover, we found that the functionalization of an interaction pair on AFM tip or substrate significantly influenced the results, implying that we must perform AFM force measurement and analyze the data with more caution.

  4. Atomic Force Microscopy and Infrared Spectroscopy Studies of Hydrogen Baked Si Surfaces

    NASA Astrophysics Data System (ADS)

    Vatel, Olivier; Verhaverbeke, Steven; Bender, Hugo; Caymax, Matty; Chollet, Frederic; Vermeire, Bert; Mertens, Paul; André, Elie; Heyns, Marc

    1993-10-01

    A H2 pre-bake at temperatures over 1050°C is typically used prior to Si epitaxial growth. In this study surface microroughness probed with tapping mode Atomic Force Microscopy (AFM) is correlated with multiple internal reflection infrared spectroscopy measurements for the different steps involved before epitaxy. A novel sample preparation technique was used for the multiple internal reflection set-up. A strong correlation was found between the presence of surface terraces and the IR double monohydride peaks for H2 annealed Si surfaces. We therefore put forward that the terraces are due to the H2 pre-bake step. These terraces remain after epitaxial deposition.

  5. Nano-scale temperature dependent visco-elastic properties of polyethylene terephthalate (PET) using atomic force microscope (AFM).

    PubMed

    Grant, Colin A; Alfouzan, Abdulrahman; Gough, Tim; Twigg, Peter C; Coates, Phil D

    2013-01-01

    Visco-elastic behaviour at the nano-level of a commonly used polymer (PET) is characterised using atomic force microscopy (AFM) at a range of temperatures. The modulus, indentation creep and relaxation time of the PET film (thickness=100 μm) is highly sensitive to temperature over an experimental temperature range of 22-175°C. The analysis showed a 40-fold increase in the amount of indentation creep on raising the temperature from 22°C to 100°C, with the most rapid rise occurring above the glass-to-rubber transition temperature (T(g)=77.1°C). At higher temperatures, close to the crystallisation temperature (T(c)=134.7°C), the indentation creep reduced to levels similar to those at temperatures below T(g). The calculated relaxation time showed a similar temperature dependence, rising from 0.6s below T(g) to 1.2s between T(g) and T(c) and falling back to 0.6s above T(c). Whereas, the recorded modulus of the thick polymer film decreases above T(g), subsequently increasing near T(c). These visco-elastic parameters are obtained via mechanical modelling of the creep curves and are correlated to the thermal phase changes that occur in PET, as revealed by differential scanning calorimetry (DSC). PMID:22750040

  6. Supramolecular Chemistry and Mechanochemistry of Macromolecules: Recent Advances by Single-Molecule Force Spectroscopy.

    PubMed

    Cheng, Bo; Cui, Shuxun

    2015-01-01

    Atomic force spectroscopy (AFM)-based single-molecule force spectroscopy (SMFS) was invented in the 1990s. Since then, SMFS has been developed into a powerful tool to study the inter- and intra-molecular interactions of macromolecules. Using SMFS, a number of problems in the field of supramolecular chemistry and mechanochemistry have been studied at the single-molecule level, which are not accessible by traditional ensemble characterization methods. In this review, the principles of SMFS are introduced, followed by the discussion of several problems of contemporary interest at the interface of supramolecular chemistry and mechanochemistry of macromolecules, including single-chain elasticity of macromolecules, interactions between water and macromolecules, interactions between macromolecules and solid surface, and the interactions in supramolecular polymers.

  7. Supramolecular Chemistry and Mechanochemistry of Macromolecules: Recent Advances by Single-Molecule Force Spectroscopy.

    PubMed

    Cheng, Bo; Cui, Shuxun

    2015-01-01

    Atomic force spectroscopy (AFM)-based single-molecule force spectroscopy (SMFS) was invented in the 1990s. Since then, SMFS has been developed into a powerful tool to study the inter- and intra-molecular interactions of macromolecules. Using SMFS, a number of problems in the field of supramolecular chemistry and mechanochemistry have been studied at the single-molecule level, which are not accessible by traditional ensemble characterization methods. In this review, the principles of SMFS are introduced, followed by the discussion of several problems of contemporary interest at the interface of supramolecular chemistry and mechanochemistry of macromolecules, including single-chain elasticity of macromolecules, interactions between water and macromolecules, interactions between macromolecules and solid surface, and the interactions in supramolecular polymers. PMID:25860255

  8. Compact metal probes: a solution for atomic force microscopy based tip-enhanced Raman spectroscopy.

    PubMed

    Rodriguez, R D; Sheremet, E; Müller, S; Gordan, O D; Villabona, A; Schulze, S; Hietschold, M; Zahn, D R T

    2012-12-01

    There are many challenges in accomplishing tip-enhanced Raman spectroscopy (TERS) and obtaining a proper tip is probably the greatest one. Since tip size, composition, and geometry are the ultimate parameters that determine enhancement of intensity and lateral resolution, the tip becomes the most critical component in a TERS experiment. However, since the discovery of TERS the cantilevers used in atomic force microscopy (AFM) have remained basically the same: commercial silicon (or silicon nitride) tips covered by a metallic coating. The main issues of using metal-coated silicon cantilevers, such as wearing off of the metal layer or increased tip radius, can be completely overcome by using all-metal cantilevers. Until now in TERS experiments such probes have only been used in a scanning tunneling microscope or in a tuning fork-based shear force microscope but not in AFM. In this work for the first time, we show the use of compact silver cantilevers that are fully compatible with contact and tapping modes in AFM demonstrating their superb performance in TERS experiments.

  9. Compact metal probes: A solution for atomic force microscopy based tip-enhanced Raman spectroscopy

    NASA Astrophysics Data System (ADS)

    Rodriguez, R. D.; Sheremet, E.; Müller, S.; Gordan, O. D.; Villabona, A.; Schulze, S.; Hietschold, M.; Zahn, D. R. T.

    2012-12-01

    There are many challenges in accomplishing tip-enhanced Raman spectroscopy (TERS) and obtaining a proper tip is probably the greatest one. Since tip size, composition, and geometry are the ultimate parameters that determine enhancement of intensity and lateral resolution, the tip becomes the most critical component in a TERS experiment. However, since the discovery of TERS the cantilevers used in atomic force microscopy (AFM) have remained basically the same: commercial silicon (or silicon nitride) tips covered by a metallic coating. The main issues of using metal-coated silicon cantilevers, such as wearing off of the metal layer or increased tip radius, can be completely overcome by using all-metal cantilevers. Until now in TERS experiments such probes have only been used in a scanning tunneling microscope or in a tuning fork-based shear force microscope but not in AFM. In this work for the first time, we show the use of compact silver cantilevers that are fully compatible with contact and tapping modes in AFM demonstrating their superb performance in TERS experiments.

  10. Capillary and van der Waals interactions on CaF2 crystals from amplitude modulation AFM force reconstruction profiles under ambient conditions.

    PubMed

    Calò, Annalisa; Robles, Oriol Vidal; Santos, Sergio; Verdaguer, Albert

    2015-01-01

    There has been much interest in the past two decades to produce experimental force profiles characteristic of the interaction between nanoscale objects or a nanoscale object and a plane. Arguably, the advent of the atomic force microscope AFM was instrumental in driving such efforts because, in principle, force profiles could be recovered directly. Nevertheless, it has taken years before techniques have developed enough as to recover the attractive part of the force with relatively low noise and without missing information on critical ranges, particularly under ambient conditions where capillary interactions are believed to dominate. Thus a systematic study of the different profiles that may arise in such situations is still lacking. Here we employ the surfaces of CaF2, on which nanoscale water films form, to report on the range and force profiles that might originate by dynamic capillary interactions occurring between an AFM tip and nanoscale water patches. Three types of force profiles were observed under ambient conditions. One in which the force decay resembles the well-known inverse-square law typical of van der Waals interactions during the first 0.5-1 nm of decay, a second one in which the force decays almost linearly, in relatively good agreement with capillary force predicted by the constant chemical potential approximation, and a third one in which the attractive force is almost constant, i.e., forms a plateau, up to 3-4 nm above the surface when the formation of a capillary neck dominates the tip-sample interaction.

  11. Probing biofouling resistant polymer brush surfaces by atomic force microscopy based force spectroscopy.

    PubMed

    Schön, Peter; Kutnyanszky, Edit; ten Donkelaar, Bas; Santonicola, M Gabriella; Tecim, Tugba; Aldred, Nick; Clare, Anthony S; Vancso, G Julius

    2013-02-01

    The protein repellency and biofouling resistance of zwitterionic poly(sulfobetaine methacrylate)(pSBMA) brushes grafted via surface initiated polymerization (SIP) from silicon and glass substrata was assessed using atomic force microscopy (AFM) adherence experiments. Laboratory settlement assays were conducted with cypris larvae of the barnacle Balanus amphitrite. AFM adherence includes the determination of contact rupture forces when AFM probe tips are withdrawn from the substratum. When the surface of the AFM tip is modified, adherence can be assessed with chemical specifity using a method known as chemical force microscopy (CFM). In this study, AFM tips were chemically functionalized with (a) fibronectin- here used as model for a nonspecifically adhering protein - and (b) arginine-glycine-aspartic acid (RGD) peptide motifs covalently attached to poly(methacrylic acid) (PMAA) brushes as biomimics of cellular adhesion receptors. Fibronectin functionalized tips showed significantly reduced nonspecific adhesion to pSBMA-modified substrata compared to bare gold (2.3±0.75 nN) and octadecanethiol (ODT) self-assembled monolayers (1.3±0.75 nN). PMAA and PMAA-RGD modified probes showed no significant adhesion to pSBMA modified silicon substrata. The results gathered through AFM protein adherence studies were complemented by laboratory fouling studies, which showed no adhesion of cypris larvae of Balanus amphitrite on pSBMA. With regard to its unusually high non-specific adsorption to a wide variety of materials the behavior of fibronectin is analogous to the barnacle cyprid temporary adhesive that also binds well to surfaces differing in polarity, charge and free energy. The antifouling efficacy of pSBMA may, therefore, be directly related to the ability of this surface to resist nonspecific protein adsorption. PMID:23138001

  12. Combined strategies for optimal detection of the contact point in AFM force-indentation curves obtained on thin samples and adherent cells.

    PubMed

    Gavara, Núria

    2016-01-01

    Atomic Force Microscopy (AFM) is a widely used tool to study cell mechanics. Current AFM setups perform high-throughput probing of living cells, generating large amounts of force-indentations curves that are subsequently analysed using a contact-mechanics model. Here we present several algorithms to detect the contact point in force-indentation curves, a crucial step to achieve fully-automated analysis of AFM-generated data. We quantify and rank the performance of our algorithms by analysing a thousand force-indentation curves obtained on thin soft homogeneous hydrogels, which mimic the stiffness and topographical profile of adherent cells. We take advantage of the fact that all the proposed algorithms are based on sequential search strategies, and show that a combination of them yields the most accurate and unbiased results. Finally, we also observe improved performance when force-indentation curves obtained on adherent cells are analysed using our combined strategy, as compared to the classical algorithm used in the majority of previous cell mechanics studies. PMID:26891762

  13. Combined strategies for optimal detection of the contact point in AFM force-indentation curves obtained on thin samples and adherent cells

    NASA Astrophysics Data System (ADS)

    Gavara, Núria

    2016-02-01

    Atomic Force Microscopy (AFM) is a widely used tool to study cell mechanics. Current AFM setups perform high-throughput probing of living cells, generating large amounts of force-indentations curves that are subsequently analysed using a contact-mechanics model. Here we present several algorithms to detect the contact point in force-indentation curves, a crucial step to achieve fully-automated analysis of AFM-generated data. We quantify and rank the performance of our algorithms by analysing a thousand force-indentation curves obtained on thin soft homogeneous hydrogels, which mimic the stiffness and topographical profile of adherent cells. We take advantage of the fact that all the proposed algorithms are based on sequential search strategies, and show that a combination of them yields the most accurate and unbiased results. Finally, we also observe improved performance when force-indentation curves obtained on adherent cells are analysed using our combined strategy, as compared to the classical algorithm used in the majority of previous cell mechanics studies.

  14. Combined strategies for optimal detection of the contact point in AFM force-indentation curves obtained on thin samples and adherent cells

    PubMed Central

    Gavara, Núria

    2016-01-01

    Atomic Force Microscopy (AFM) is a widely used tool to study cell mechanics. Current AFM setups perform high-throughput probing of living cells, generating large amounts of force-indentations curves that are subsequently analysed using a contact-mechanics model. Here we present several algorithms to detect the contact point in force-indentation curves, a crucial step to achieve fully-automated analysis of AFM-generated data. We quantify and rank the performance of our algorithms by analysing a thousand force-indentation curves obtained on thin soft homogeneous hydrogels, which mimic the stiffness and topographical profile of adherent cells. We take advantage of the fact that all the proposed algorithms are based on sequential search strategies, and show that a combination of them yields the most accurate and unbiased results. Finally, we also observe improved performance when force-indentation curves obtained on adherent cells are analysed using our combined strategy, as compared to the classical algorithm used in the majority of previous cell mechanics studies. PMID:26891762

  15. Force spectroscopy of membrane hardness of SH-SY5Y neuroblastoma cells before and after differentiation

    NASA Astrophysics Data System (ADS)

    Kwon, Sangwoo; Yang, Woochul; Choi, Yun Kyong; Park, Jung Keuck

    2014-05-01

    Atomic force microscopy (AFM) is utilized in many studies for measuring the structure and the physical characteristics of soft and bio materials. In particular, the force spectroscopy function in the AFM system allows us to explore the mechanical properties of bio cells. In this study, we probe the variation in the membrane hardness of human neuroblastoma SH-SY5Y cells (SH-cells) before and after differentiation by using force spectroscopy. The SH-cell, which is usually differentiated by using a chemical treatment with retinoic acid (RA), is a neuronal cell line employed widely as an in-vitro model for neuroscience research. In force spectroscopy, the force-distance curves are obtained from both the original and the RA-treated cells while the AFM tip approaches and pushes on the cell membranes. The slope deduced from linear region in the force-distance curve is the spring constant and corresponds to the hardness of the cell membrane. The spring constant of the RA-treated cells (0.597 ± 0.010 nN/nm) was smaller than that of the original cells (0.794 ± 0.010 nN/nm), reflecting a hardness decrease in the cells differentiated with the RA treatments. The results clearly demonstrated that the differentiated cells are softer than the original cells. The change in the elasticity of the differentiated cells might be caused by morphological modification during differentiation process. We suggest that force spectroscopy can be employed as a novel method to determine the degree of differentiation of stem cells into various functional cells.

  16. Quadrature phase interferometer for high resolution force spectroscopy

    SciTech Connect

    Paolino, Pierdomenico; Aguilar Sandoval, Felipe A.; Bellon, Ludovic

    2013-09-15

    In this article, we present a deflection measurement setup for Atomic Force Microscopy (AFM). It is based on a quadrature phase differential interferometer: we measure the optical path difference between a laser beam reflecting above the cantilever tip and a reference beam reflecting on the static base of the sensor. A design with very low environmental susceptibility and another allowing calibrated measurements on a wide spectral range are described. Both enable a very high resolution (down to 2.5×10{sup −15} m/√(Hz)), illustrated by thermal noise measurements on AFM cantilevers. They present an excellent long-term stability and a constant sensitivity independent of the optical phase of the interferometer. A quick review shows that our precision is equaling or out-performing the best results reported in the literature, but for a much larger deflection range, up to a few μm.

  17. Qplus AFM driven nanostencil.

    PubMed

    Grévin, B; Fakir, M; Hayton, J; Brun, M; Demadrille, R; Faure-Vincent, J

    2011-06-01

    We describe the development of a novel setup, in which large stencils with suspended silicon nitride membranes are combined with atomic force microscopy (AFM) regulation by using tuning forks. This system offers the possibility to perform separate AFM and nanostencil operations, as well as combined modes when using stencil chips with integrated tips. The flexibility and performances are demonstrated through a series of examples, including wide AFM scans in closed loop mode, probe positioning repeatability of a few tens of nanometer, simultaneous evaporation of large (several hundred of micron square) and nanoscopic metals and fullerene patterns in static, multistep, and dynamic modes. This approach paves the way for further developments, as it fully combines the advantages of conventional stenciling with the ones of an AFM driven shadow mask. PMID:21721701

  18. Leukocyte-endothelium interaction: measurement by laser tweezers force spectroscopy.

    PubMed

    Wang, Shi-Kang; Chiu, Jeng-Jiann; Lee, Ming-Rou; Chou, Shih-Chin; Chen, Li-Jing; Hwang, Ned H C

    2006-09-01

    Leukocyte adhesion to vascular endothelium is an initial step of many inflammatory diseases. Although the atomic force microscopy (AFM) measurements of leukocyte-endothelial interaction have been recently introduced. with cell adhesion force unbinding curves (CAFUC). We obtained pico-Newton force in the initial interaction between a single living THP-1 cell and HUVEC monolayer using a custom-built laser tweezers (LT) system. The measured quantities included the non-linear force-distance relationship, and the effect of yielding in cell detachment. It is possible to introduce a time scale into the LT cell-detachment experiments for further exploration and more detailed information on the viscoelastic properties of living cells. PMID:16960761

  19. Capillary and van der Waals interactions on CaF2 crystals from amplitude modulation AFM force reconstruction profiles under ambient conditions

    PubMed Central

    Calò, Annalisa; Robles, Oriol Vidal; Santos, Sergio

    2015-01-01

    Summary There has been much interest in the past two decades to produce experimental force profiles characteristic of the interaction between nanoscale objects or a nanoscale object and a plane. Arguably, the advent of the atomic force microscope AFM was instrumental in driving such efforts because, in principle, force profiles could be recovered directly. Nevertheless, it has taken years before techniques have developed enough as to recover the attractive part of the force with relatively low noise and without missing information on critical ranges, particularly under ambient conditions where capillary interactions are believed to dominate. Thus a systematic study of the different profiles that may arise in such situations is still lacking. Here we employ the surfaces of CaF2, on which nanoscale water films form, to report on the range and force profiles that might originate by dynamic capillary interactions occurring between an AFM tip and nanoscale water patches. Three types of force profiles were observed under ambient conditions. One in which the force decay resembles the well-known inverse-square law typical of van der Waals interactions during the first 0.5–1 nm of decay, a second one in which the force decays almost linearly, in relatively good agreement with capillary force predicted by the constant chemical potential approximation, and a third one in which the attractive force is almost constant, i.e., forms a plateau, up to 3–4 nm above the surface when the formation of a capillary neck dominates the tip–sample interaction. PMID:25977852

  20. The ReactorAFM: non-contact atomic force microscope operating under high-pressure and high-temperature catalytic conditions.

    PubMed

    Roobol, S B; Cañas-Ventura, M E; Bergman, M; van Spronsen, M A; Onderwaater, W G; van der Tuijn, P C; Koehler, R; Ofitserov, A; van Baarle, G J C; Frenken, J W M

    2015-03-01

    An Atomic Force Microscope (AFM) has been integrated in a miniature high-pressure flow reactor for in-situ observations of heterogeneous catalytic reactions under conditions similar to those of industrial processes. The AFM can image model catalysts such as those consisting of metal nanoparticles on flat oxide supports in a gas atmosphere up to 6 bar and at a temperature up to 600 K, while the catalytic activity can be measured using mass spectrometry. The high-pressure reactor is placed inside an Ultrahigh Vacuum (UHV) system to supplement it with standard UHV sample preparation and characterization techniques. To demonstrate that this instrument successfully bridges both the pressure gap and the materials gap, images have been recorded of supported palladium nanoparticles catalyzing the oxidation of carbon monoxide under high-pressure, high-temperature conditions.

  1. The ReactorAFM: Non-contact atomic force microscope operating under high-pressure and high-temperature catalytic conditions

    SciTech Connect

    Roobol, S. B.; Cañas-Ventura, M. E.; Bergman, M.; Spronsen, M. A. van; Onderwaater, W. G.; Tuijn, P. C. van der; Koehler, R.; Frenken, J. W. M.; Ofitserov, A.; Baarle, G. J. C. van

    2015-03-15

    An Atomic Force Microscope (AFM) has been integrated in a miniature high-pressure flow reactor for in-situ observations of heterogeneous catalytic reactions under conditions similar to those of industrial processes. The AFM can image model catalysts such as those consisting of metal nanoparticles on flat oxide supports in a gas atmosphere up to 6 bar and at a temperature up to 600 K, while the catalytic activity can be measured using mass spectrometry. The high-pressure reactor is placed inside an Ultrahigh Vacuum (UHV) system to supplement it with standard UHV sample preparation and characterization techniques. To demonstrate that this instrument successfully bridges both the pressure gap and the materials gap, images have been recorded of supported palladium nanoparticles catalyzing the oxidation of carbon monoxide under high-pressure, high-temperature conditions.

  2. Physical properties of poly(lactic-co-glycolic) and poly(ethylene glycol) nanoparticles for drug delivery using atomic force microscopy (AFM) and electrostatic nanolithography

    NASA Astrophysics Data System (ADS)

    Lyuksyutov, Sergei; Fedin, Igor; Nedashkivska, Victoria; Lyuksyutova, Caterina; Geldenhuys, Werner; Sutariya, Vijay

    2010-03-01

    Nanoparticles (NP) of biodegradable polymers poly(lactic-co-glycolic)(PLGA) and poly(ethylene glycol) (PEG) are potential drug delivery components for biomedical applications. The NP based on PLGA or PEG can be directed to accumulate in cancer tumor cells with the use of anti-bodies which are conjugated to the NP. The NP's size distribution is the critical property for biochemical affinity and therefore delivery to the specific target organs. We used an atomic force microscopy (AFM) to characterize the NP size and AFM electrostatic nanolithography (AFMEN) to study the behavior of PEG-PLGA NP under the extreme electric fields exceeding 10^9 V m-1. AFMEN allows the displacement of molecules along the lines of the electric field due to electrostatic polarization. This study has an important practical application for the optimum design of NP with the correct characteristics for drug delivery.

  3. The origin of the "snap-in" in the force curve between AFM probe and the water/gas interface of nanobubbles.

    PubMed

    Song, Yang; Zhao, Binyu; Zhang, Lijuan; Lü, Junhong; Wang, Shuo; Dong, Yaming; Hu, Jun

    2014-02-24

    The long-range attractive force or "snap-in" is an important phenomenon usually occurring when a solid particle interacts with a water/gas interface. By using PeakForce quantitative nanomechanics the origin of snap-in in the force curve between the atomic force microscopy (AFM) probe and the water/gas interface of nanobubbles has been investigated. The snap-in frequently happened when the probe was preserved for a certain time or after being used for imaging solid surfaces under atmospheric conditions. In contrast, imaging in liquids rarely induced a snap-in. After a series of control experiments, it was found that the snap-in can be attributed to hydrophobic interactions between the water/gas interface and the AFM probe, which was either modified or contaminated with hydrophobic material. The hydrophobic contamination could be efficiently removed by a conventional plasma-cleaning treatment, which prevents the occurring of the snap-in. In addition, the adsorption of sodium dodecyl sulfate onto the nanobubble surface changed the water/gas interface into hydrophilic, which also eliminated the snap-in phenomenon.

  4. Versatile method for AFM-tip functionalization with biomolecules: fishing a ligand by means of an in situ click reaction.

    PubMed

    Kumar, Rakesh; Ramakrishna, Shivaprakash N; Naik, Vikrant V; Chu, Zonglin; Drew, Michael E; Spencer, Nicholas D; Yamakoshi, Yoko

    2015-04-21

    A facile and universal method for the functionalization of an AFM tip has been developed for chemical force spectroscopy (CFS) studies of intermolecular interactions of biomolecules. A click reaction between tripod-acetylene and an azide-linker-ligand molecule was successfully carried out on the AFM tip surface and used for the CFS study of ligand-receptor interactions.

  5. Charge injection in thin dielectric layers by atomic force microscopy: influence of geometry and material work function of the AFM tip on the injection process.

    PubMed

    Villeneuve-Faure, C; Makasheva, K; Boudou, L; Teyssedre, G

    2016-06-17

    Charge injection and retention in thin dielectric layers remain critical issues for the reliability of many electronic devices because of their association with a large number of failure mechanisms. To overcome this drawback, a deep understanding of the mechanisms leading to charge injection close to the injection area is needed. Even though the charge injection is extensively studied and reported in the literature to characterize the charge storage capability of dielectric materials, questions about charge injection mechanisms when using atomic force microscopy (AFM) remain open. In this paper, a thorough study of charge injection by using AFM in thin plasma-processed amorphous silicon oxynitride layers with properties close to that of thermal silica layers is presented. The study considers the impact of applied voltage polarity, work function of the AFM tip coating and tip curvature radius. A simple theoretical model was developed and used to analyze the obtained experimental results. The electric field distribution is computed as a function of tip geometry. The obtained experimental results highlight that after injection in the dielectric layer the charge lateral spreading is mainly controlled by the radial electric field component independently of the carrier polarity. The injected charge density is influenced by the nature of electrode metal coating (work function) and its geometry (tip curvature radius). The electron injection is mainly ruled by the Schottky injection barrier through the field electron emission mechanism enhanced by thermionic electron emission. The hole injection mechanism seems to differ from the electron one depending on the work function of the metal coating. Based on the performed analysis, it is suggested that for hole injection by AFM, pinning of the metal Fermi level with the metal-induced gap states in the studied silicon oxynitride layers starts playing a role in the injection mechanisms. PMID:27158768

  6. Charge injection in thin dielectric layers by atomic force microscopy: influence of geometry and material work function of the AFM tip on the injection process

    NASA Astrophysics Data System (ADS)

    Villeneuve-Faure, C.; Makasheva, K.; Boudou, L.; Teyssedre, G.

    2016-06-01

    Charge injection and retention in thin dielectric layers remain critical issues for the reliability of many electronic devices because of their association with a large number of failure mechanisms. To overcome this drawback, a deep understanding of the mechanisms leading to charge injection close to the injection area is needed. Even though the charge injection is extensively studied and reported in the literature to characterize the charge storage capability of dielectric materials, questions about charge injection mechanisms when using atomic force microscopy (AFM) remain open. In this paper, a thorough study of charge injection by using AFM in thin plasma-processed amorphous silicon oxynitride layers with properties close to that of thermal silica layers is presented. The study considers the impact of applied voltage polarity, work function of the AFM tip coating and tip curvature radius. A simple theoretical model was developed and used to analyze the obtained experimental results. The electric field distribution is computed as a function of tip geometry. The obtained experimental results highlight that after injection in the dielectric layer the charge lateral spreading is mainly controlled by the radial electric field component independently of the carrier polarity. The injected charge density is influenced by the nature of electrode metal coating (work function) and its geometry (tip curvature radius). The electron injection is mainly ruled by the Schottky injection barrier through the field electron emission mechanism enhanced by thermionic electron emission. The hole injection mechanism seems to differ from the electron one depending on the work function of the metal coating. Based on the performed analysis, it is suggested that for hole injection by AFM, pinning of the metal Fermi level with the metal-induced gap states in the studied silicon oxynitride layers starts playing a role in the injection mechanisms.

  7. Charge injection in thin dielectric layers by atomic force microscopy: influence of geometry and material work function of the AFM tip on the injection process.

    PubMed

    Villeneuve-Faure, C; Makasheva, K; Boudou, L; Teyssedre, G

    2016-06-17

    Charge injection and retention in thin dielectric layers remain critical issues for the reliability of many electronic devices because of their association with a large number of failure mechanisms. To overcome this drawback, a deep understanding of the mechanisms leading to charge injection close to the injection area is needed. Even though the charge injection is extensively studied and reported in the literature to characterize the charge storage capability of dielectric materials, questions about charge injection mechanisms when using atomic force microscopy (AFM) remain open. In this paper, a thorough study of charge injection by using AFM in thin plasma-processed amorphous silicon oxynitride layers with properties close to that of thermal silica layers is presented. The study considers the impact of applied voltage polarity, work function of the AFM tip coating and tip curvature radius. A simple theoretical model was developed and used to analyze the obtained experimental results. The electric field distribution is computed as a function of tip geometry. The obtained experimental results highlight that after injection in the dielectric layer the charge lateral spreading is mainly controlled by the radial electric field component independently of the carrier polarity. The injected charge density is influenced by the nature of electrode metal coating (work function) and its geometry (tip curvature radius). The electron injection is mainly ruled by the Schottky injection barrier through the field electron emission mechanism enhanced by thermionic electron emission. The hole injection mechanism seems to differ from the electron one depending on the work function of the metal coating. Based on the performed analysis, it is suggested that for hole injection by AFM, pinning of the metal Fermi level with the metal-induced gap states in the studied silicon oxynitride layers starts playing a role in the injection mechanisms.

  8. Kinetic Ductility and Force-Spike Resistance of Proteins from Single-Molecule Force Spectroscopy.

    PubMed

    Cossio, Pilar; Hummer, Gerhard; Szabo, Attila

    2016-08-23

    Ductile materials can absorb spikes in mechanical force, whereas brittle ones fail catastrophically. Here we develop a theory to quantify the kinetic ductility of single molecules from force spectroscopy experiments, relating force-spike resistance to the differential responses of the intact protein and the unfolding transition state to an applied mechanical force. We introduce a class of unistable one-dimensional potential surfaces that encompass previous models as special cases and continuously cover the entire range from ductile to brittle. Compact analytic expressions for force-dependent rates and rupture-force distributions allow us to analyze force-clamp and force-ramp pulling experiments. We find that the force-transmitting protein domains of filamin and titin are kinetically ductile when pulled from their two termini, making them resistant to force spikes. For the mechanostable muscle protein titin, a highly ductile model reconciles data over 10 orders of magnitude in force loading rate from experiment and simulation.

  9. Kinetic Ductility and Force-Spike Resistance of Proteins from Single-Molecule Force Spectroscopy.

    PubMed

    Cossio, Pilar; Hummer, Gerhard; Szabo, Attila

    2016-08-23

    Ductile materials can absorb spikes in mechanical force, whereas brittle ones fail catastrophically. Here we develop a theory to quantify the kinetic ductility of single molecules from force spectroscopy experiments, relating force-spike resistance to the differential responses of the intact protein and the unfolding transition state to an applied mechanical force. We introduce a class of unistable one-dimensional potential surfaces that encompass previous models as special cases and continuously cover the entire range from ductile to brittle. Compact analytic expressions for force-dependent rates and rupture-force distributions allow us to analyze force-clamp and force-ramp pulling experiments. We find that the force-transmitting protein domains of filamin and titin are kinetically ductile when pulled from their two termini, making them resistant to force spikes. For the mechanostable muscle protein titin, a highly ductile model reconciles data over 10 orders of magnitude in force loading rate from experiment and simulation. PMID:27558726

  10. Simulation of Force Spectroscopy Experiments on Galacturonic Acid Oligomers

    PubMed Central

    Cybulska, Justyna; Brzyska, Agnieszka; Zdunek, Artur; Woliński, Krzysztof

    2014-01-01

    Pectins, forming a matrix for cellulose and hemicellulose, determine the mechanics of plant cell walls. They undergo salient structural changes during their development. In the presence of divalent cations, usually calcium, pectins can form gel-like structures. Because of their importance they have been the subject of many force spectroscopy experiments, which have examined the conformational changes and molecular tensions due to external forces. The most abundant unit present in the pectin backbone is polygalacturonic acid. Unfortunately, experimental force spectroscopy on polygalacturonic acid molecules is still not a trivial task. The mechanism of the single-molecule response to external forces can be inferred by theoretical methods. Therefore, in this work we simulated such force spectroscopy experiments using the Enforced Geometry Optimization (EGO) method. We examined the oligomeric (up to hexamer) structures of α-D-galacturonic acid exposed to external stretching forces. The EGO simulation of the force spectroscopy appropriately reproduced the experimental course of the enforced conformational transition: chair →inverted chair via the twisted boat conformation(s) in the pyranose ring of α-D-galacturonic acid. Additionally, our theoretical approach also allowed to determine the minimum oligomer size adequate for the description of nano-mechanical properties of (poly)-α-D-galacturonic acid. PMID:25229407

  11. Oxidative dissolution of chalcopyrite by Acidithiobacillus ferrooxidans analyzed by electrochemical impedance spectroscopy and atomic force microscopy.

    PubMed

    Bevilaqua, D; Diéz-Perez, I; Fugivara, C S; Sanz, F; Benedetti, A V; Garcia, O

    2004-08-01

    The microbiological leaching of chalcopyrite (CuFeS(2)) is of great interest because of its potential application to many CuFeS(2)-rich ore materials. However, the efficiency of the microbiological process is very limited because this mineral is one of the most refractory to bacterial attack. Knowledge of bacterial role during chalcopyrite oxidation is very important in order to improve the efficiency of bioleaching operation. The oxidative dissolution of a massive chalcopyrite electrode by Acidithiobacillus ferrooxidans was evaluated by electrochemical impedance spectroscopy (EIS) and atomic force microscopy (AFM). A massive chalcopyrite electrode was utilized in a Tait-type electrochemical cell in acid medium for different immersion times in the presence or absence of bacterium. The differences observed in the impedance diagrams were correlated with the adhesion process of bacteria on the mineral surface. PMID:15219250

  12. Laser irradiation effects on the CdTe/ZnTe quantum dot structure studied by Raman and AFM spectroscopy

    SciTech Connect

    Zielony, E.; Placzek-Popko, E.; Henrykowski, A.; Gumienny, Z.; Kamyczek, P.; Jacak, J.; Nowakowski, P.; Karczewski, G.

    2012-09-15

    Micro-Raman spectroscopy has been applied to investigate the impact of laser irradiation on semiconducting CdTe/ZnTe quantum dots (QDs) structures. A reference sample (without dots) was also studied for comparison. Both samples were grown by molecular beam epitaxy technique on the p-type GaAs substrate. The Raman spectra have been recorded for different time of a laser exposure and for various laser powers. The spectra for both samples exhibit peak related to the localized longitudinal (LO) ZnTe phonon of a wavenumber equal to 210 cm{sup -1}. For the QD sample, a broad band corresponding to the LO CdTe phonon related to the QD-layer appears at a wavenumber of 160 cm{sup -1}. With increasing time of a laser beam exposure and laser power, the spectra get dominated by tellurium-related peaks appearing at wavenumbers around 120 cm{sup -1} and 140 cm{sup -1}. Simultaneously, the ZnTe surface undergoes rising damage, with the formation of Te aggregates at the pinhole edge as reveal atomic force microscopy observations. Local temperature of irradiated region has been estimated from the anti-Stokes/Stokes ratio of the Te modes intensity and it was found to be close or exceeding ZnTe melting point. Thus, the laser damage can be explained by the ablation process.

  13. Single-Cell Force Spectroscopy of Probiotic Bacteria

    PubMed Central

    Beaussart, Audrey; El-Kirat-Chatel, Sofiane; Herman, Philippe; Alsteens, David; Mahillon, Jacques; Hols, Pascal; Dufrêne, Yves F.

    2013-01-01

    Single-cell force spectroscopy is a powerful atomic force microscopy modality in which a single living cell is attached to the atomic force microscopy cantilever to quantify the forces that drive cell-cell and cell-substrate interactions. Although various single-cell force spectroscopy protocols are well established for animal cells, application of the method to individual bacterial cells remains challenging, mainly owing to the lack of appropriate methods for the controlled attachment of single live cells on cantilevers. We present a nondestructive protocol for single-bacterial cell force spectroscopy, which combines the use of colloidal probe cantilevers and of a bioinspired polydopamine wet adhesive. Living cells from the probiotic species Lactobacillus plantarum are picked up with a polydopamine-coated colloidal probe, enabling us to quantify the adhesion forces between single bacteria and biotic (lectin monolayer) or abiotic (hydrophobic monolayer) surfaces. These minimally invasive single-cell experiments provide novel, to our knowledge, insight into the specific and nonspecific forces driving the adhesion of L. plantarum, and represent a generic platform for studying the molecular mechanisms of cell adhesion in probiotic and pathogenic bacteria. PMID:23663831

  14. On artifacts in single-molecule force spectroscopy

    PubMed Central

    Cossio, Pilar; Hummer, Gerhard; Szabo, Attila

    2015-01-01

    In typical force spectroscopy experiments, a small biomolecule is attached to a soft polymer linker that is pulled with a relatively large bead or cantilever. At constant force, the total extension stochastically changes between two (or more) values, indicating that the biomolecule undergoes transitions between two (or several) conformational states. In this paper, we consider the influence of the dynamics of the linker and mesoscopic pulling device on the force-dependent rate of the conformational transition extracted from the time dependence of the total extension, and the distribution of rupture forces in force-clamp and force-ramp experiments, respectively. For these different experiments, we derive analytic expressions for the observables that account for the mechanical response and dynamics of the pulling device and linker. Possible artifacts arise when the characteristic times of the pulling device and linker become comparable to, or slower than, the lifetimes of the metastable conformational states, and when the highly anharmonic regime of stretched linkers is probed at high forces. We also revisit the problem of relating force-clamp and force-ramp experiments, and identify a linker and loading rate-dependent correction to the rates extracted from the latter. The theory provides a framework for both the design and the quantitative analysis of force spectroscopy experiments by highlighting, and correcting for, factors that complicate their interpretation. PMID:26540730

  15. Probing local bias-induced transitions using photothermal excitation contact resonance atomic force microscopy and voltage spectroscopy

    DOE PAGES

    Li, Qian; Jesse, Stephen; Tselev, Alexander; Collins, Liam; Yu, Pu; Kravchenko, Ivan; Kalinin, Sergei V.; Balke, Nina

    2015-01-05

    In this paper, nanomechanical properties are closely related to the states of matter, including chemical composition, crystal structure, mesoscopic domain configuration, etc. Investigation of these properties at the nanoscale requires not only static imaging methods, e.g., contact resonance atomic force microscopy (CR-AFM), but also spectroscopic methods capable of revealing their dependence on various external stimuli. Here we demonstrate the voltage spectroscopy of CR-AFM, which was realized by combining photothermal excitation (as opposed to the conventional piezoacoustic excitation method) with the band excitation technique. We applied this spectroscopy to explore local bias-induced phenomena ranging from purely physical to surface electromechanical andmore » electrochemical processes. Our measurements show that the changes in the surface properties associated with these bias-induced transitions can be accurately assessed in a fast and dynamic manner, using resonance frequency as a signature. Finally, with many of the advantages offered by photothermal excitation, contact resonance voltage spectroscopy not only is expected to find applications in a broader field of nanoscience but also will provide a basis for future development of other nanoscale elastic spectroscopies.« less

  16. Probing local bias-induced transitions using photothermal excitation contact resonance atomic force microscopy and voltage spectroscopy

    SciTech Connect

    Li, Qian; Jesse, Stephen; Tselev, Alexander; Collins, Liam; Yu, Pu; Kravchenko, Ivan; Kalinin, Sergei V.; Balke, Nina

    2015-01-05

    In this paper, nanomechanical properties are closely related to the states of matter, including chemical composition, crystal structure, mesoscopic domain configuration, etc. Investigation of these properties at the nanoscale requires not only static imaging methods, e.g., contact resonance atomic force microscopy (CR-AFM), but also spectroscopic methods capable of revealing their dependence on various external stimuli. Here we demonstrate the voltage spectroscopy of CR-AFM, which was realized by combining photothermal excitation (as opposed to the conventional piezoacoustic excitation method) with the band excitation technique. We applied this spectroscopy to explore local bias-induced phenomena ranging from purely physical to surface electromechanical and electrochemical processes. Our measurements show that the changes in the surface properties associated with these bias-induced transitions can be accurately assessed in a fast and dynamic manner, using resonance frequency as a signature. Finally, with many of the advantages offered by photothermal excitation, contact resonance voltage spectroscopy not only is expected to find applications in a broader field of nanoscience but also will provide a basis for future development of other nanoscale elastic spectroscopies.

  17. Absorption Spectroscopy and Imaging from the Visible through Mid-IR with 20 nm Resolution Using AFM probes

    NASA Astrophysics Data System (ADS)

    Centrone, Andrea

    2015-03-01

    Correlated nanoscale composition and optical property maps are important to engineer nanomaterials in applications ranging from photovoltaics to sensing and therapeutics. Wavelengths (λs) from the visible to near-IR probe electronic transitions in materials, providing information regarding band gap and defects while light in mid-IR probes vibrational transitions and provide chemical composition. However, light diffraction limits the lateral resolution of conventional micro-spectroscopic techniques to approximately λ/2, which is insufficient to image nanomaterials. Additionally, the λ-dependent resolution impedes direct comparison of spectral maps from different spectral ranges. Photo Thermal Induced Resonance (PTIR) is a novel technique that circumvents light diffraction by employing an AFM tip as a local detector for measuring light absorption with λ-independent nanoscale resolution. Our PTIR setup combines an AFM microscope with three lasers providing λ-tunability from 500 nm to 16000 nm continuously. The AFM tip transduces locally the sample thermal expansion induced by light absorption into large cantilever oscillations. Local absorption spectra (electronic or vibrational) and maps are obtained recording the amplitude of the tip deflection as a function of λ and position, respectively. The working principles of the PTIR technique will be described first, and nano-patterned polymer samples will be used to evaluate its lateral resolution, sensitivity and linearity. Results show that the PTIR signal intensity is proportional to the local absorbed energy suggesting applicability of this technique for quantitative chemical analysis at nanoscale, at least for thin (less than 1000 nm thick) samples. Additionally, a λ-independent resolution as high as 20 nm is demonstrated across the whole spectral range. In the second part of the talk, PTIR will be applied to image the dark plasmonic resonance of gold Asymmetric Split Ring Resonators (A-SRRs) in the mid

  18. Subfemtonewton Force Spectroscopy at the Thermal Limit in Liquids

    NASA Astrophysics Data System (ADS)

    Liu, Lulu; Kheifets, Simon; Ginis, Vincent; Capasso, Federico

    2016-06-01

    We demonstrate thermally limited force spectroscopy using a probe formed by a dielectric microsphere optically trapped in water near a dielectric surface. We achieve force resolution below 1 fN in 100 s, corresponding to a 2 Å rms displacement of the probe. Our measurement combines a calibrated evanescent wave particle tracking technique and a lock-in detection method. We demonstrate the accuracy of our method by measurement of the height-dependent force exerted on the probe by an evanescent wave, the results of which are in agreement with Mie theory calculations.

  19. Local tunneling decay length and Kelvin probe force spectroscopy

    NASA Astrophysics Data System (ADS)

    Albrecht, Florian; Fleischmann, Martin; Scheer, Manfred; Gross, Leo; Repp, Jascha

    2015-12-01

    In the past, current-distance spectroscopy has been widely applied to determine variations of the work function at surfaces. While for homogeneous sample areas this technique is commonly accepted to yield at least qualitative results, its applicability to atomic-scale variations has not been proven neither right nor wrong. Here we benchmark measurements of the current-distance decay constant against the well established Kelvin probe force spectroscopy for four distinctly different cases with atomic-scale variations of the local contact potential. The two techniques yield quite different results. Whereas the maps of the current-distance decay constant are consistent with being topographical artifacts, the Kelvin probe force spectroscopy maps show variations of the local contact potential difference in agreement with expected surface dipoles. This comparison clarifies that maps of the current-distance decay constant are not suited to directly characterize contact potential variations at surfaces on atomic length scales.

  20. Reconstructing Folding Energy Landscapes by Single-Molecule Force Spectroscopy

    PubMed Central

    Woodside, Michael T.; Block, Steven M.

    2015-01-01

    Folding may be described conceptually in terms of trajectories over a landscape of free energies corresponding to different molecular configurations. In practice, energy landscapes can be difficult to measure. Single-molecule force spectroscopy (SMFS), whereby structural changes are monitored in molecules subjected to controlled forces, has emerged as a powerful tool for probing energy landscapes. We summarize methods for reconstructing landscapes from force spectroscopy measurements under both equilibrium and nonequilibrium conditions. Other complementary, but technically less demanding, methods provide a model-dependent characterization of key features of the landscape. Once reconstructed, energy landscapes can be used to study critical folding parameters, such as the characteristic transition times required for structural changes and the effective diffusion coefficient setting the timescale for motions over the landscape. We also discuss issues that complicate measurement and interpretation, including the possibility of multiple states or pathways and the effects of projecting multiple dimensions onto a single coordinate. PMID:24895850

  1. Reconstructing folding energy landscapes by single-molecule force spectroscopy.

    PubMed

    Woodside, Michael T; Block, Steven M

    2014-01-01

    Folding may be described conceptually in terms of trajectories over a landscape of free energies corresponding to different molecular configurations. In practice, energy landscapes can be difficult to measure. Single-molecule force spectroscopy (SMFS), whereby structural changes are monitored in molecules subjected to controlled forces, has emerged as a powerful tool for probing energy landscapes. We summarize methods for reconstructing landscapes from force spectroscopy measurements under both equilibrium and nonequilibrium conditions. Other complementary, but technically less demanding, methods provide a model-dependent characterization of key features of the landscape. Once reconstructed, energy landscapes can be used to study critical folding parameters, such as the characteristic transition times required for structural changes and the effective diffusion coefficient setting the timescale for motions over the landscape. We also discuss issues that complicate measurement and interpretation, including the possibility of multiple states or pathways and the effects of projecting multiple dimensions onto a single coordinate.

  2. Study of the interactions between endolysin and bacterial peptidoglycan on S. aureus by dynamic force spectroscopy

    NASA Astrophysics Data System (ADS)

    Liu, Jianli; Zhang, Xuejie; Yang, Hang; Yuan, Jinghe; Wei, Hongping; Yu, Junping; Fang, Xiaohong

    2015-09-01

    The cell wall binding domain (CBD) of bacteriophage lysins can recognize target bacteria with extraordinary specificity through binding to bacterial peptidoglycan, thus it is a promising new probe to identify the corresponding bacterial pathogen. In this work, we used atomic force microscopy (AFM) based single-molecule force spectroscopy to investigate the interaction between the CBD of lysin PlyV12 (PlyV12C) and pathogenic bacterium Staphylococcus aureus (S. aureus). The binding forces of PlyV12C with S. aureus have been measured, and the dissociation process of their binding complex has been characterized. Furthermore, we compared the interactions of PlyV12C-S. aureus and antibody-S. aureus. It is revealed that PlyV12C has a comparable affinity to bacterial peptidoglycans as that of the S. aureus antibody. The results provide new information on the binding properties of lysin CBD with bacterium, and the application of lysin CBD in bacterium detection.

  3. Resolving dual binding conformations of cellulosome cohesin-dockerin complexes using single-molecule force spectroscopy

    PubMed Central

    Jobst, Markus A; Milles, Lukas F; Schoeler, Constantin; Ott, Wolfgang; Fried, Daniel B; Bayer, Edward A; Gaub, Hermann E; Nash, Michael A

    2015-01-01

    Receptor-ligand pairs are ordinarily thought to interact through a lock and key mechanism, where a unique molecular conformation is formed upon binding. Contrary to this paradigm, cellulosomal cohesin-dockerin (Coh-Doc) pairs are believed to interact through redundant dual binding modes consisting of two distinct conformations. Here, we combined site-directed mutagenesis and single-molecule force spectroscopy (SMFS) to study the unbinding of Coh:Doc complexes under force. We designed Doc mutations to knock out each binding mode, and compared their single-molecule unfolding patterns as they were dissociated from Coh using an atomic force microscope (AFM) cantilever. Although average bulk measurements were unable to resolve the differences in Doc binding modes due to the similarity of the interactions, with a single-molecule method we were able to discriminate the two modes based on distinct differences in their mechanical properties. We conclude that under native conditions wild-type Doc from Clostridium thermocellum exocellulase Cel48S populates both binding modes with similar probabilities. Given the vast number of Doc domains with predicteddual binding modes across multiple bacterial species, our approach opens up newpossibilities for understanding assembly and catalytic properties of a broadrange of multi-enzyme complexes. DOI: http://dx.doi.org/10.7554/eLife.10319.001 PMID:26519733

  4. Force dependency of biochemical reactions measured by single-molecule force-clamp spectroscopy.

    PubMed

    Popa, Ionel; Kosuri, Pallav; Alegre-Cebollada, Jorge; Garcia-Manyes, Sergi; Fernandez, Julio M

    2013-01-01

    Here we describe a protocol for using force-clamp spectroscopy to precisely quantify the effect of force on biochemical reactions. A calibrated force is used to control the exposure of reactive sites in a single polyprotein substrate composed of repeated domains. The use of polyproteins allows the identification of successful single-molecule recordings from unambiguous mechanical unfolding fingerprints. Biochemical reactions are then measured directly by detecting the length changes of the substrate held at a constant force. We present the layout of a force-clamp spectrometer along with protocols to design and conduct experiments. These experiments measure reaction kinetics as a function of applied force. We show sample data of the force dependency of two different reactions, protein unfolding and disulfide reduction. These data, which can be acquired in just a few days, reveal mechanistic details of the reactions that currently cannot be resolved by any other technique.

  5. Force dependency of biochemical reactions measured by single molecule force-clamp spectroscopy

    PubMed Central

    Popa, Ionel; Kosuri, Pallav; Alegre-Cebollada, Jorge; Garcia-Manyes, Sergi; Fernandez, Julio M.

    2015-01-01

    Here we describe a protocol for using force-clamp spectroscopy to precisely quantify the effect of force on biochemical reactions. A calibrated force is used to control the exposure of reactive sites in a single polyprotein substrate composed of repeated domains. The use of polyproteins allows the identification of successful single-molecule recordings from unambiguous mechanical unfolding fingerprints. Biochemical reactions are then measured directly by detecting the length changes of the substrate held at a constant force. We present the layout of a force-clamp spectrometer along with protocols to design and conduct experiments. These experiments measure reaction kinetics as a function of applied force. We show sample data of the force dependency of two different reactions, protein unfolding and disulfide reduction. These data, which can be acquired in just a few days, reveal mechanistic details of the reactions that currently cannot be resolved by any other technique. PMID:23744288

  6. Probing Cytoskeletal Structures by Coupling Optical Superresolution and AFM Techniques for a Correlative Approach

    PubMed Central

    Chacko, Jenu Varghese; Zanacchi, Francesca Cella; Diaspro, Alberto

    2013-01-01

    In this article, we describe and show the application of some of the most advanced fluorescence superresolution techniques, STED AFM and STORM AFM microscopy towards imaging of cytoskeletal structures, such as microtubule filaments. Mechanical and structural properties can play a relevant role in the investigation of cytoskeletal structures of interest, such as microtubules, that provide support to the cell structure. In fact, the mechanical properties, such as the local stiffness and the elasticity, can be investigated by AFM force spectroscopy with tens of nanometers resolution. Force curves can be analyzed in order to obtain the local elasticity (and the Young's modulus calculation by fitting the force curves from every pixel of interest), and the combination with STED/STORM microscopy integrates the measurement with high specificity and yields superresolution structural information. This hybrid modality of superresolution-AFM working is a clear example of correlative multimodal microscopy. PMID:24027190

  7. Vibrational analysis of single-layered piezoelectric AFM microcantilever in amplitude mode by considering the capillary force

    NASA Astrophysics Data System (ADS)

    Habibnejad Korayem, Alireza; Habibnejad Korayem, Moharam; Ghaderi, Reza

    2014-12-01

    In this article, the vibrational behavior of a microcantilever (MC) with an extended piezoelectric layer in the air ambient undergoes examination. To model the vibrational motion of this type of cantilever, the Hamilton's principle has been used. For this purpose, the MC vibrational equation has been derived by the assumption of the continuous beam based on the Euler-Bernoulli beam theory. By adopting the finite element method (FEM), the MC differential equation has been solved. In the present simulation not only van der Waals and contact forces but also the capillary forces resulting from the condensation of the water vapors in air on MC tip have been considered. The results illustrate that the force between the sample surface and the probe affects the MC amplitude; furthermore, it causes the reduction in the resonance frequency. In addition, to reduce the time delay during topography from the surface roughness, it is better to select MCs with larger width and length and smaller thickness. Furthermore, the results indicate that the best imaging takes place when the vibration is in its second vibrational mode. Finally, the effects of MC geometric parameters on the time delay between the starting moment of surface roughness and the moment of variation in the MC amplitude (surface roughness topography) have been analyzed.

  8. Probing the interaction of individual amino acids with inorganic surfaces using atomic force spectroscopy.

    PubMed

    Razvag, Yair; Gutkin, Vitaly; Reches, Meital

    2013-08-13

    This article describes single-molecule force spectroscopy measurements of the interaction between individual amino acid residues and inorganic surfaces in an aqueous solution. In each measurement, there is an amino acid residue, lysine, glutamate, phenylalanine, leucine, or glutamine, and each represents a class of amino acids (positively or negatively charged, aromatic, nonpolar, and polar). Force-distance curves measured the interaction of the individual amino acid bound to a silicon atomic force microscope (AFM) tip with a silcon substrate, cut from a single-crystal wafer, or mica. Using this method, we were able to measure low adhesion forces (below 300 pN) and could clearly determine the strength of interactions between the individual amino acid residues and the inorganic substrate. In addition, we observed how changes in the pH and ionic strength of the solution affected the adsorption of the residues to the substrates. Our results pinpoint the important role of hydrophobic interactions among the amino acids and the substrate, where hydrophobic phenylalanine exhibited the strongest adhesion to a silicon substrate. Additionally, electrostatic interactions also contributed to the adsorption of amino acid residues to inorganic substrates. A change in the pH or ionic strength values of the buffer altered the strength of interactions among the amino acids and the substrate. We concluded that the interplay between the hydrophobic forces and electrostatic interactions will determine the strength of adsorption among the amino acids and the surface. Overall, these results contribute to our understanding of the interaction at the organic-inorganic interface. These results may have implications for our perception of the specificity of peptide binding to inorganic surfaces. Consequently, it would possibly lead to a better design of composite materials and devices.

  9. Single-cell force spectroscopy of pili-mediated adhesion

    NASA Astrophysics Data System (ADS)

    Sullan, Ruby May A.; Beaussart, Audrey; Tripathi, Prachi; Derclaye, Sylvie; El-Kirat-Chatel, Sofiane; Li, James K.; Schneider, Yves-Jacques; Vanderleyden, Jos; Lebeer, Sarah; Dufrêne, Yves F.

    2013-12-01

    Although bacterial pili are known to mediate cell adhesion to a variety of substrates, the molecular interactions behind this process are poorly understood. We report the direct measurement of the forces guiding pili-mediated adhesion, focusing on the medically important probiotic bacterium Lactobacillus rhamnosus GG (LGG). Using non-invasive single-cell force spectroscopy (SCFS), we quantify the adhesion forces between individual bacteria and biotic (mucin, intestinal cells) or abiotic (hydrophobic monolayers) surfaces. On hydrophobic surfaces, bacterial pili strengthen adhesion through remarkable nanospring properties, which - presumably - enable the bacteria to resist high shear forces under physiological conditions. On mucin, nanosprings are more frequent and adhesion forces larger, reflecting the influence of specific pili-mucin bonds. Interestingly, these mechanical responses are no longer observed on human intestinal Caco-2 cells. Rather, force curves exhibit constant force plateaus with extended ruptures reflecting the extraction of membrane nanotethers. These single-cell analyses provide novel insights into the molecular mechanisms by which piliated bacteria colonize surfaces (nanosprings, nanotethers), and offer exciting avenues in nanomedicine for understanding and controlling the adhesion of microbial cells (probiotics, pathogens).

  10. Quantitatively Resolving Multivalent Interactions on Macroscopic Scale Using Force Spectroscopy

    PubMed Central

    Hu, Qiongzheng; Yang, Haopeng; Wang, Yuhong; Xu, Shoujun

    2016-01-01

    Multivalent interactions remain difficult to be characterized and consequently controlled, particularly on a macroscopic scale. Using force-induced remnant magnetization spectroscopy (FIRMS), we have resolved the single-, double-, and triple- biotin—streptavidin interactions, multivalent DNA interactions and CXCL12-CXCR4 interactions, on millimetre-scale surfaces. Our results establish FIRMS as a viable method for systematic resolution and controlled formation of multivalent interactions. PMID:26864087

  11. Force spectroscopy of biomolecular folding and binding: theory meets experiment

    NASA Astrophysics Data System (ADS)

    Dudko, Olga

    2015-03-01

    Conformational transitions in biological macromolecules usually serve as the mechanism that brings biomolecules into their working shape and enables their biological function. Single-molecule force spectroscopy probes conformational transitions by applying force to individual macromolecules and recording their response, or ``mechanical fingerprints,'' in the form of force-extension curves. However, how can we decode these fingerprints so that they reveal the kinetic barriers and the associated timescales of a biological process? I will present an analytical theory of the mechanical fingerprints of macromolecules. The theory is suitable for decoding such fingerprints to extract the barriers and timescales. The application of the theory will be illustrated through recent studies on protein-DNA interactions and the receptor-ligand complexes involved in blood clot formation.

  12. Direct measurements of the interaction between pyrene and graphite in aqueous media by single molecule force spectroscopy: understanding the pi-pi interactions.

    PubMed

    Zhang, Yiheng; Liu, Chuanjun; Shi, Weiqing; Wang, Zhiqiang; Dai, Liming; Zhang, Xi

    2007-07-17

    Pyrene derivatives can absorb onto the surface of carbon nanotubes and graphite particles through pi-pi interactions to functionalize these inorganic building blocks with organic surface moieties. Using single molecule force spectroscopy, we have demonstrated the first direct measurement of the interaction between pyrene and a graphite surface. In particular, we have connected a pyrene molecule onto an AFM tip via a flexible poly(ethylene glycol) (PEG) chain to ensure the formation of a molecular bridge. The pi-pi interaction between pyrene and graphite is thus indicated to be approximately 55 pN with no hysteresis between the desorption and adhesion forces. PMID:17590031

  13. Discriminating Intercalative Effects of Threading Intercalator Nogalamycin, from Classical Intercalator Daunomycin, Using Single Molecule Atomic Force Spectroscopy

    PubMed Central

    Sett, S.; Ghosh, S.; Rakshit, T.; Mukhopadhyay, R.

    2016-01-01

    DNA threading intercalators are a unique class of intercalating agents, albeit little biophysical information is available on their intercalative actions. Herein, the intercalative effects of nogalamycin, which is a naturally-occurring DNA threading intercalator, have been investigated by high-resolution atomic force microscopy (AFM) and spectroscopy (AFS). The results have been compared with those of the well-known chemotherapeutic drug daunomycin, which is a non-threading classical intercalator bearing structural similarity to nogalamycin. A comparative AFM assessment revealed a greater increase in DNA contour length over the entire incubation period of 48 h for nogalamycin treatment, whereas the contour length increase manifested faster in case of daunomycin. The elastic response of single DNA molecules to an externally applied force was investigated by the single molecule AFS approach. Characteristic mechanical fingerprints in the overstretching behaviour clearly distinguished the nogalamycin/daunomycin-treated dsDNA from untreated dsDNA—the former appearing less elastic than the latter, and the nogalamycin-treated DNA distinguished from the daunomycin-treated DNA—the classically intercalated dsDNA appearing the least elastic. A single molecule AFS-based discrimination of threading intercalation from the classical type is being reported for the first time. PMID:27183010

  14. An integrated instrumental setup for the combination of atomic force microscopy with optical spectroscopy.

    PubMed

    Owen, R J; Heyes, C D; Knebel, D; Röcker, C; Nienhaus, G U

    2006-07-01

    In recent years, the study of single biomolecules using fluorescence microscopy and atomic force microscopy (AFM) techniques has resulted in a plethora of new information regarding the physics underlying these complex biological systems. It is especially advantageous to be able to measure the optical, topographical, and mechanical properties of single molecules simultaneously. Here an AFM is used that is especially designed for integration with an inverted optical microscope and that has a near-infrared light source (850 nm) to eliminate interference between the optical experiment and the AFM operation. The Tip Assisted Optics (TAO) system consists of an additional 100 x 100-microm(2) X-Y scanner for the sample, which can be independently and simultaneously used with the AFM scanner. This allows the offset to be removed between the confocal optical image obtained with the sample scanner and the simultaneously acquired AFM topography image. The tip can be positioned exactly into the optical focus while the user can still navigate within the AFM image for imaging or manipulation of the sample. Thus the tip-enhancement effect can be maximized and it becomes possible to perform single molecule manipulation experiments within the focus of a confocal optical image. Here this is applied to simultaneous measurement of single quantum dot fluorescence and topography with high spatial resolution.

  15. Scanning near-field optical/atomic force microscopy for fluorescence imaging and spectroscopy of biomaterials in air and liquid: Observation of recombinant Escherichia coli with gene coding to green fluorescent protein

    NASA Astrophysics Data System (ADS)

    Muramatsu, Hiroshi; Chiba, Norio; Ataka, Tatsuaki; Iwabuchi, Shinichiro; Nagatani, Naoki; Tamiya, Eiichi; Fujihira, Masamichi

    1996-11-01

    We have developed a system of scanning near-field optical/atomic force microscopy (SNOM/AFM) for fluorescence imaging and spectroscopy of biomaterials in air and liquid. SNOM/AFM uses a bent optical fiber simultaneously as a dynamic force AFM cantilever and a SNOM probe. Optical resolution of SNOM images shows about 50 nm in an illumination mode for a standard sample of a patterned chromium layer of 20 nm thickness on a quartz glass plate. The SNOM/AFM system contains a photon counting system and polychrometer/ICCD (intensified charge coupled device) system for observation of the fluorescence image and spectrograph of micro areas, respectively. The gene coding to green fluorescence protein (GFP) was cloned in recombinant Escherichia coli (E. coli). Topography, fluorescence image and spectrograph of recombinant E. coli by SNOM/AFM showed a difference in fluorescence in individual E. coli. Fluorescence activity of GFP can thus be used as a convenient indicator of transformation. SNOM/AFM is also applicable to observe immobilized E. coli on a glass plate in water with a liquid chamber and may allow the viewing of observation of floating organisms.

  16. Nonlinearly Additive Forces in Multivalent Ligand Binding to a Single Protein Revealed with Force Spectroscopy

    SciTech Connect

    Ratto, T V; Rudd, R E; Langry, K C; Balhorn, R L; McElfresh, M W

    2005-07-15

    We present evidence of multivalent interactions between a single protein molecule and multiple carbohydrates at a pH where the protein can bind four ligands. The evidence is based not only on measurements of the force required to rupture the bonds formed between ConcanavalinA (ConA) and {alpha}-D-mannose, but also on an analysis of the polymer-extension force curves to infer the polymer architecture that binds the protein to the cantilever and the ligands to the substrate. We find that although the rupture forces for multiple carbohydrate connections to a single protein are larger than the rupture force for a single connection, they do not scale additively with increasing number. Specifically, the most common rupture forces are approximately 46, 66, and 85 pN, which we argue corresponds to 1, 2, and 3 ligands being pulled simultaneously from a single protein as corroborated by an analysis of the linkage architecture. As in our previous work polymer tethers allow us to discriminate between specific and non-specific binding. We analyze the binding configuration (i.e. serial versus parallel connections) through fitting the polymer stretching data with modified Worm-Like Chain (WLC) models that predict how the effective stiffness of the tethers is affected by multiple connections. This analysis establishes that the forces we measure are due to single proteins interacting with multiple ligands, the first force spectroscopy study that establishes single-molecule multivalent binding unambiguously.

  17. Characterization of novel sufraces by FTIR spectroscopy and atomic force microscopy for food pathogen detection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Single molecular detection of pathogens and toxins of interest to food safety is within grasp using technology such as Atomic Force Microscopy. Using antibodies or specific aptamers connected to the AFM tip make it possible to detect a pathogen molecule on a surface. However, it also becomes necess...

  18. High-resolution high-speed dynamic mechanical spectroscopy of cells and other soft materials with the help of atomic force microscopy.

    PubMed

    Dokukin, M; Sokolov, I

    2015-07-28

    Dynamic mechanical spectroscopy (DMS), which allows measuring frequency-dependent viscoelastic properties, is important to study soft materials, tissues, biomaterials, polymers. However, the existing DMS techniques (nanoindentation) have limited resolution when used on soft materials, preventing them from being used to study mechanics at the nanoscale. The nanoindenters are not capable of measuring cells, nanointerfaces of composite materials. Here we present a highly accurate DMS modality, which is a combination of three different methods: quantitative nanoindentation (nanoDMA), gentle force and fast response of atomic force microscopy (AFM), and Fourier transform (FT) spectroscopy. This new spectroscopy (which we suggest to call FT-nanoDMA) is fast and sensitive enough to allow DMS imaging of nanointerfaces, single cells, while attaining about 100x improvements on polymers in both spatial (to 10-70 nm) and temporal resolution (to 0.7 s/pixel) compared to the current art. Multiple frequencies are measured simultaneously. The use of 10 frequencies are demonstrated here (up to 300 Hz which is a rather relevant range for biological materials and polymers, in both ambient conditions and liquid). The method is quantitatively verified on known polymers and demonstrated on cells and polymers blends. Analysis shows that FT-nanoDMA is highly quantitative. The FT-nanoDMA spectroscopy can easily be implemented in the existing AFMs.

  19. High-resolution high-speed dynamic mechanical spectroscopy of cells and other soft materials with the help of atomic force microscopy

    PubMed Central

    Dokukin, M.; Sokolov, I.

    2015-01-01

    Dynamic mechanical spectroscopy (DMS), which allows measuring frequency-dependent viscoelastic properties, is important to study soft materials, tissues, biomaterials, polymers. However, the existing DMS techniques (nanoindentation) have limited resolution when used on soft materials, preventing them from being used to study mechanics at the nanoscale. The nanoindenters are not capable of measuring cells, nanointerfaces of composite materials. Here we present a highly accurate DMS modality, which is a combination of three different methods: quantitative nanoindentation (nanoDMA), gentle force and fast response of atomic force microscopy (AFM), and Fourier transform (FT) spectroscopy. This new spectroscopy (which we suggest to call FT-nanoDMA) is fast and sensitive enough to allow DMS imaging of nanointerfaces, single cells, while attaining about 100x improvements on polymers in both spatial (to 10–70 nm) and temporal resolution (to 0.7s/pixel) compared to the current art. Multiple frequencies are measured simultaneously. The use of 10 frequencies are demonstrated here (up to 300 Hz which is a rather relevant range for biological materials and polymers, in both ambient conditions and liquid). The method is quantitatively verified on known polymers and demonstrated on cells and polymers blends. Analysis shows that FT-nanoDMA is highly quantitative. The FT-nanoDMA spectroscopy can easily be implemented in the existing AFMs. PMID:26218346

  20. Towards force spectroscopy of single tip-link bonds

    NASA Astrophysics Data System (ADS)

    Koussa, Mounir A.; Sotomayor, Marcos; Wong, Wesley P.; Corey, David P.

    2015-12-01

    Inner-ear mechanotransduction relies on tip links, fine protein filaments made of cadherin-23 and protocadherin-15 that convey tension to mechanosensitive channels at the tips of hair-cell stereocilia. The tip-link cadherins are thought to form a heterotetrameric complex, with two cadherin-23 molecules forming the upper part of the filament and two protocadherin-15 molecules forming the lower end. The interaction between cadherin-23 and protocadherin-15 is mediated by their N-terminal tips. Missense mutations that modify the interaction interface impair binding and lead to deafness. Molecular dynamics simulations predict that the tip-link bond is mechanically strong enough to withstand forces in hair cells, but its experimentally determined strength is unknown. We have developed molecular tools to facilitate single-molecule force spectroscopy on the tip link bond. Self-assembling DNA nanoswitches are functionalized with the interacting tips of cadherin-23 and protocadherin-15 using the enzyme sortase under conditions that preserve protein function. These tip link nanoswitches are designed to provide a signature force-extension profile. This molecular signature should allow us to identify single-molecule rupture events in pulling experiments.

  1. Fundamental High-Speed Limits in Single-Molecule, Single-Cell, and Nanoscale Force Spectroscopies.

    PubMed

    Amo, Carlos A; Garcia, Ricardo

    2016-07-26

    Force spectroscopy is enhancing our understanding of single-biomolecule, single-cell, and nanoscale mechanics. Force spectroscopy postulates the proportionality between the interaction force and the instantaneous probe deflection. By studying the probe dynamics, we demonstrate that the total force acting on the probe has three different components: the interaction, the hydrodynamic, and the inertial. The amplitudes of those components depend on the ratio between the resonant frequency and the frequency at which the data are measured. A force-distance curve provides a faithful measurement of the interaction force between two molecules when the inertial and hydrodynamic components are negligible. Otherwise, force spectroscopy measurements will underestimate the value of unbinding forces. Neglecting the above force components requires the use of frequency ratios in the 50-500 range. These ratios will limit the use of high-speed methods in force spectroscopy. The theory is supported by numerical simulations. PMID:27359243

  2. Fundamental High-Speed Limits in Single-Molecule, Single-Cell, and Nanoscale Force Spectroscopies.

    PubMed

    Amo, Carlos A; Garcia, Ricardo

    2016-07-26

    Force spectroscopy is enhancing our understanding of single-biomolecule, single-cell, and nanoscale mechanics. Force spectroscopy postulates the proportionality between the interaction force and the instantaneous probe deflection. By studying the probe dynamics, we demonstrate that the total force acting on the probe has three different components: the interaction, the hydrodynamic, and the inertial. The amplitudes of those components depend on the ratio between the resonant frequency and the frequency at which the data are measured. A force-distance curve provides a faithful measurement of the interaction force between two molecules when the inertial and hydrodynamic components are negligible. Otherwise, force spectroscopy measurements will underestimate the value of unbinding forces. Neglecting the above force components requires the use of frequency ratios in the 50-500 range. These ratios will limit the use of high-speed methods in force spectroscopy. The theory is supported by numerical simulations.

  3. Charge-state dynamics in electrostatic force spectroscopy

    NASA Astrophysics Data System (ADS)

    Ondráček, Martin; Hapala, Prokop; Jelínek, Pavel

    2016-07-01

    We present a numerical model that allows us to study the response of an oscillating probe in electrostatic force spectroscopy to charge switching in quantum dots at various time scales. The model provides more insight into the behavior of frequency shift and dissipated energy under different scanning conditions when measuring a temporarily charged quantum dot on a surface. Namely, we analyze the dependence of the frequency shift, the dissipated energy, and their fluctuations on the resonance frequency of the tip and on the electron tunneling rates across the tip-quantum dot and quantum dot-sample junctions. We discuss two complementary approaches to simulating the charge dynamics, a stochastic and a deterministic one. In addition, we derive analytic formulas valid for small amplitudes, describing relations between the frequency shift, dissipated energy, and the characteristic rates driving the charging and discharging processes.

  4. Subnanometre enzyme mechanics probed by single-molecule force spectroscopy

    PubMed Central

    Pelz, Benjamin; Žoldák, Gabriel; Zeller, Fabian; Zacharias, Martin; Rief, Matthias

    2016-01-01

    Enzymes are molecular machines that bind substrates specifically, provide an adequate chemical environment for catalysis and exchange products rapidly, to ensure fast turnover rates. Direct information about the energetics that drive conformational changes is difficult to obtain. We used subnanometre single-molecule force spectroscopy to study the energetic drive of substrate-dependent lid closing in the enzyme adenylate kinase. Here we show that in the presence of the bisubstrate inhibitor diadenosine pentaphosphate (AP5A), closing and opening of both lids is cooperative and tightly coupled to inhibitor binding. Surprisingly, binding of the substrates ADP and ATP exhibits a much smaller energetic drive towards the fully closed state. Instead, we observe a new dominant energetic minimum with both lids half closed. Our results, combining experiment and molecular dynamics simulations, give detailed mechanical insights into how an enzyme can cope with the seemingly contradictory requirements of rapid substrate exchange and tight closing, to ensure efficient catalysis. PMID:26906294

  5. Subnanometre enzyme mechanics probed by single-molecule force spectroscopy

    NASA Astrophysics Data System (ADS)

    Pelz, Benjamin; Žoldák, Gabriel; Zeller, Fabian; Zacharias, Martin; Rief, Matthias

    2016-02-01

    Enzymes are molecular machines that bind substrates specifically, provide an adequate chemical environment for catalysis and exchange products rapidly, to ensure fast turnover rates. Direct information about the energetics that drive conformational changes is difficult to obtain. We used subnanometre single-molecule force spectroscopy to study the energetic drive of substrate-dependent lid closing in the enzyme adenylate kinase. Here we show that in the presence of the bisubstrate inhibitor diadenosine pentaphosphate (AP5A), closing and opening of both lids is cooperative and tightly coupled to inhibitor binding. Surprisingly, binding of the substrates ADP and ATP exhibits a much smaller energetic drive towards the fully closed state. Instead, we observe a new dominant energetic minimum with both lids half closed. Our results, combining experiment and molecular dynamics simulations, give detailed mechanical insights into how an enzyme can cope with the seemingly contradictory requirements of rapid substrate exchange and tight closing, to ensure efficient catalysis.

  6. Subnanometre enzyme mechanics probed by single-molecule force spectroscopy.

    PubMed

    Pelz, Benjamin; Žoldák, Gabriel; Zeller, Fabian; Zacharias, Martin; Rief, Matthias

    2016-02-24

    Enzymes are molecular machines that bind substrates specifically, provide an adequate chemical environment for catalysis and exchange products rapidly, to ensure fast turnover rates. Direct information about the energetics that drive conformational changes is difficult to obtain. We used subnanometre single-molecule force spectroscopy to study the energetic drive of substrate-dependent lid closing in the enzyme adenylate kinase. Here we show that in the presence of the bisubstrate inhibitor diadenosine pentaphosphate (AP5A), closing and opening of both lids is cooperative and tightly coupled to inhibitor binding. Surprisingly, binding of the substrates ADP and ATP exhibits a much smaller energetic drive towards the fully closed state. Instead, we observe a new dominant energetic minimum with both lids half closed. Our results, combining experiment and molecular dynamics simulations, give detailed mechanical insights into how an enzyme can cope with the seemingly contradictory requirements of rapid substrate exchange and tight closing, to ensure efficient catalysis.

  7. Exploring the folding pattern of a polymer chain in a single crystal by combining single-molecule force spectroscopy and steered molecular dynamics simulations.

    PubMed

    Song, Yu; Feng, Wei; Liu, Kai; Yang, Peng; Zhang, Wenke; Zhang, Xi

    2013-03-26

    Understanding the folding pattern of a single polymer chain within its single crystal will shed light on the mechanism of crystallization. Here, we use the combined techniques of atomic force microscopy (AFM)-based single-molecule force spectroscopy (SMFS) and steered molecular dynamics (SMD) simulations to study the folding pattern of a polyethylene oxide (PEO) chain in its single crystal. Our results show that the folding pattern of a PEO chain in the crystal formed in dilute solution follows the adjacent re-entry folding model. While in the crystal obtained from the melt, the nonadjacent folding with large and irregular loops contributes to big force fluctuations in the force-extension curves. The method established here can offer a novel strategy to directly unravel the chain-folding pattern of polymer single crystals at single-molecule level.

  8. Fundamental High-Speed Limits in Single-Molecule, Single-Cell, and Nanoscale Force Spectroscopies

    PubMed Central

    2016-01-01

    Force spectroscopy is enhancing our understanding of single-biomolecule, single-cell, and nanoscale mechanics. Force spectroscopy postulates the proportionality between the interaction force and the instantaneous probe deflection. By studying the probe dynamics, we demonstrate that the total force acting on the probe has three different components: the interaction, the hydrodynamic, and the inertial. The amplitudes of those components depend on the ratio between the resonant frequency and the frequency at which the data are measured. A force–distance curve provides a faithful measurement of the interaction force between two molecules when the inertial and hydrodynamic components are negligible. Otherwise, force spectroscopy measurements will underestimate the value of unbinding forces. Neglecting the above force components requires the use of frequency ratios in the 50–500 range. These ratios will limit the use of high-speed methods in force spectroscopy. The theory is supported by numerical simulations. PMID:27359243

  9. Characterization of chemically and enzymatically treated hemp fibres using atomic force microscopy and spectroscopy

    NASA Astrophysics Data System (ADS)

    George, Michael; Mussone, Paolo G.; Abboud, Zeinab; Bressler, David C.

    2014-09-01

    The mechanical and moisture resistance properties of natural fibre reinforced composites are dependent on the adhesion between the matrix of choice and the fibre. The main goal of this study was to investigate the effect of NaOH swelling of hemp fibres prior to enzymatic treatment and a novel chemical sulfonic acid method on the physical properties of hemp fibres. The colloidal properties of treated hemp fibres were studied exclusively using an atomic force microscope. AFM imaging in tapping mode revealed that each treatment rendered the surface topography of the hemp fibres clean and exposed the individual fibre bundles. Hemp fibres treated with laccase had no effect on the surface adhesion forces measured. Interestingly, mercerization prior to xylanase + cellulase and laccase treatments resulted in greater enzyme access evident in the increased adhesion force measurements. Hemp fibres treated with sulfonic acid showed an increase in surface de-fibrillation and smoothness. A decrease in adhesion forces for 4-aminotoulene-3-sulfonic acid (AT3S) treated fibres suggested a reduction in surface polarity. This work demonstrated that AFM can be used as a tool to estimate the surface forces and roughness for modified fibres and that enzymatic coupled with chemical methods can be used to improve the surface properties of natural fibres for composite applications. Further, this work is one of the first that offers some insight into the effect of mercerization prior to enzymes and the effect on the surface topography. AFM will be used to selectively screen treated fibres for composite applications based on the adhesion forces associated with the colloidal interface between the AFM tip and the fibre surfaces.

  10. An open source/real-time atomic force microscope architecture to perform customizable force spectroscopy experiments.

    PubMed

    Materassi, Donatello; Baschieri, Paolo; Tiribilli, Bruno; Zuccheri, Giampaolo; Samorì, Bruno

    2009-08-01

    We describe the realization of an atomic force microscope architecture designed to perform customizable experiments in a flexible and automatic way. Novel technological contributions are given by the software implementation platform (RTAI-LINUX), which is free and open source, and from a functional point of view, by the implementation of hard real-time control algorithms. Some other technical solutions such as a new way to estimate the optical lever constant are described as well. The adoption of this architecture provides many degrees of freedom in the device behavior and, furthermore, allows one to obtain a flexible experimental instrument at a relatively low cost. In particular, we show how such a system has been employed to obtain measures in sophisticated single-molecule force spectroscopy experiments [Fernandez and Li, Science 303, 1674 (2004)]. Experimental results on proteins already studied using the same methodologies are provided in order to show the reliability of the measure system.

  11. Segmented nanofibers of spider dragline silk: atomic force microscopy and single-molecule force spectroscopy.

    PubMed

    Oroudjev, E; Soares, J; Arcdiacono, S; Thompson, J B; Fossey, S A; Hansma, H G

    2002-04-30

    Despite its remarkable materials properties, the structure of spider dragline silk has remained unsolved. Results from two probe microscopy techniques provide new insights into the structure of spider dragline silk. A soluble synthetic protein from dragline silk spontaneously forms nanofibers, as observed by atomic force microscopy. These nanofibers have a segmented substructure. The segment length and amino acid sequence are consistent with a slab-like shape for individual silk protein molecules. The height and width of nanofiber segments suggest a stacking pattern of slab-like molecules in each nanofiber segment. This stacking pattern produces nano-crystals in an amorphous matrix, as observed previously by NMR and x-ray diffraction of spider dragline silk. The possible importance of nanofiber formation to native silk production is discussed. Force spectra for single molecules of the silk protein demonstrate that this protein unfolds through a number of rupture events, indicating a modular substructure within single silk protein molecules. A minimal unfolding module size is estimated to be around 14 nm, which corresponds to the extended length of a single repeated module, 38 amino acids long. The structure of this spider silk protein is distinctly different from the structures of other proteins that have been analyzed by single-molecule force spectroscopy, and the force spectra show correspondingly novel features.

  12. Probing the PEDOT:PSS/cell interface with conductive colloidal probe AFM-SECM

    NASA Astrophysics Data System (ADS)

    Knittel, P.; Zhang, H.; Kranz, C.; Wallace, G. G.; Higgins, M. J.

    2016-02-01

    Conductive colloidal probe Atomic Force-Scanning Electrochemical Microscopy (AFM-SECM) is a new approach, which employs electrically insulated AFM probes except for a gold-coated colloid located at the end of the cantilever. Hence, force measurements can be performed while biasing the conductive colloid under physiological conditions. Moreover, such colloids can be modified by electrochemical polymerization resulting, e.g. in conductive polymer-coated spheres, which in addition may be loaded with specific dopants. In contrast to other AFM-based single cell force spectroscopy measurements, these probes allow adhesion measurements at the cell-biomaterial interface on multiple cells in a rapid manner while the properties of the polymer can be changed by applying a bias. In addition, spatially resolved electrochemical information e.g., oxygen reduction can be obtained simultaneously. Conductive colloid AFM-SECM probes modified with poly(3,4-ethylenedioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS) are used for single cell force measurements in mouse fibroblasts and single cell interactions are investigated as a function of the applied potential.Conductive colloidal probe Atomic Force-Scanning Electrochemical Microscopy (AFM-SECM) is a new approach, which employs electrically insulated AFM probes except for a gold-coated colloid located at the end of the cantilever. Hence, force measurements can be performed while biasing the conductive colloid under physiological conditions. Moreover, such colloids can be modified by electrochemical polymerization resulting, e.g. in conductive polymer-coated spheres, which in addition may be loaded with specific dopants. In contrast to other AFM-based single cell force spectroscopy measurements, these probes allow adhesion measurements at the cell-biomaterial interface on multiple cells in a rapid manner while the properties of the polymer can be changed by applying a bias. In addition, spatially resolved electrochemical

  13. Mismatch detection in DNA monolayers by atomic force microscopy and electrochemical impedance spectroscopy

    PubMed Central

    Ambrosetti, Elena; Scoles, Giacinto; Casalis, Loredana

    2016-01-01

    Summary Background: DNA hybridization is at the basis of most current technologies for genotyping and sequencing, due to the unique properties of DNA base-pairing that guarantee a high grade of selectivity. Nonetheless the presence of single base mismatches or not perfectly matched sequences can affect the response of the devices and the major challenge is, nowadays, to distinguish a mismatch of a single base and, at the same time, unequivocally differentiate devices read-out of fully and partially matching sequences. Results: We present here two platforms based on different sensing strategies, to detect mismatched and/or perfectly matched complementary DNA strands hybridization into ssDNA oligonucleotide monolayers. The first platform exploits atomic force microscopy-based nanolithography to create ssDNA nano-arrays on gold surfaces. AFM topography measurements then monitor the variation of height of the nanostructures upon biorecognition and then follow annealing at different temperatures. This strategy allowed us to clearly detect the presence of mismatches. The second strategy exploits the change in capacitance at the interface between an ssDNA-functionalized gold electrode and the solution due to the hybridization process in a miniaturized electrochemical cell. Through electrochemical impedance spectroscopy measurements on extended ssDNA self-assembled monolayers we followed in real-time the variation of capacitance, being able to distinguish, through the difference in hybridization kinetics, not only the presence of single, double or triple mismatches in the complementary sequence, but also the position of the mismatched base pair with respect to the electrode surface. Conclusion: We demonstrate here two platforms based on different sensing strategies as sensitive and selective tools to discriminate mismatches. Our assays are ready for parallelization and can be used in the detection and quantification of single nucleotide mismatches in microRNAs or in

  14. Custom AFM for X-ray beamlines: in situ biological investigations under physiological conditions.

    PubMed

    Gumí-Audenis, B; Carlà, F; Vitorino, M V; Panzarella, A; Porcar, L; Boilot, M; Guerber, S; Bernard, P; Rodrigues, M S; Sanz, F; Giannotti, M I; Costa, L

    2015-11-01

    A fast atomic force microscope (AFM) has been developed that can be installed as a sample holder for grazing-incidence X-ray experiments at solid/gas or solid/liquid interfaces. It allows a wide range of possible investigations, including soft and biological samples under physiological conditions (hydrated specimens). The structural information obtained using the X-rays is combined with the data gathered with the AFM (morphology and mechanical properties), providing a unique characterization of the specimen and its dynamics in situ during an experiment. In this work, lipid monolayers and bilayers in air or liquid environment have been investigated by means of AFM, both with imaging and force spectroscopy, and X-ray reflectivity. In addition, this combination allows the radiation damage induced by the beam on the sample to be studied, as has been observed on DOPC and DPPC supported lipid bilayers under physiological conditions. PMID:26524300

  15. Custom AFM for X-ray beamlines: in situ biological investigations under physiological conditions

    PubMed Central

    Gumí-Audenis, B.; Carlà, F.; Vitorino, M. V.; Panzarella, A.; Porcar, L.; Boilot, M.; Guerber, S.; Bernard, P.; Rodrigues, M. S.; Sanz, F.; Giannotti, M. I.; Costa, L.

    2015-01-01

    A fast atomic force microscope (AFM) has been developed that can be installed as a sample holder for grazing-incidence X-ray experiments at solid/gas or solid/liquid interfaces. It allows a wide range of possible investigations, including soft and biological samples under physiological conditions (hydrated specimens). The structural information obtained using the X-rays is combined with the data gathered with the AFM (morphology and mechanical properties), providing a unique characterization of the specimen and its dynamics in situ during an experiment. In this work, lipid monolayers and bilayers in air or liquid environment have been investigated by means of AFM, both with imaging and force spectroscopy, and X-ray reflectivity. In addition, this combination allows the radiation damage induced by the beam on the sample to be studied, as has been observed on DOPC and DPPC supported lipid bilayers under physiological conditions. PMID:26524300

  16. Quantifying Molecular-Level Cell Adhesion on Electroactive Conducting Polymers using Electrochemical-Single Cell Force Spectroscopy

    PubMed Central

    Zhang, Hongrui; Molino, Paul J.; Wallace, Gordon G.; Higgins, Michael J.

    2015-01-01

    Single Cell Force Spectroscopy was combined with Electrochemical-AFM to quantify the adhesion between live single cells and conducting polymers whilst simultaneously applying a voltage to electrically switch the polymer from oxidized to reduced states. The cell-conducting polymer adhesion represents the non-specific interaction between cell surface glycocalyx molecules and polymer groups such as sulfonate and dodecylbenzene groups, which rearrange their orientation during electrical switching. Single cell adhesion significantly increases as the polymer is switched from an oxidized to fully reduced state, indicating stronger cell binding to sulfonate groups as opposed to hydrophobic groups. This increase in single cell adhesion is concomitant with an increase in surface hydrophilicity and uptake of cell media, driven by cation movement, into the polymer film during electrochemical reduction. Binding forces between the glycocalyx and polymer surface are indicative of molecular-level interactions and during electrical stimulation there is a decrease in both the binding force and stiffness of the adhesive bonds. The study provides insight into the effects of electrochemical switching on cell adhesion at the cell-conducting polymer interface and is more broadly applicable to elucidating the binding of cell adhesion molecules in the presence of electrical fields and directly at electrode interfaces. PMID:26335299

  17. Quantifying Molecular-Level Cell Adhesion on Electroactive Conducting Polymers using Electrochemical-Single Cell Force Spectroscopy

    NASA Astrophysics Data System (ADS)

    Zhang, Hongrui; Molino, Paul J.; Wallace, Gordon G.; Higgins, Michael J.

    2015-09-01

    Single Cell Force Spectroscopy was combined with Electrochemical-AFM to quantify the adhesion between live single cells and conducting polymers whilst simultaneously applying a voltage to electrically switch the polymer from oxidized to reduced states. The cell-conducting polymer adhesion represents the non-specific interaction between cell surface glycocalyx molecules and polymer groups such as sulfonate and dodecylbenzene groups, which rearrange their orientation during electrical switching. Single cell adhesion significantly increases as the polymer is switched from an oxidized to fully reduced state, indicating stronger cell binding to sulfonate groups as opposed to hydrophobic groups. This increase in single cell adhesion is concomitant with an increase in surface hydrophilicity and uptake of cell media, driven by cation movement, into the polymer film during electrochemical reduction. Binding forces between the glycocalyx and polymer surface are indicative of molecular-level interactions and during electrical stimulation there is a decrease in both the binding force and stiffness of the adhesive bonds. The study provides insight into the effects of electrochemical switching on cell adhesion at the cell-conducting polymer interface and is more broadly applicable to elucidating the binding of cell adhesion molecules in the presence of electrical fields and directly at electrode interfaces.

  18. The Study of Biomolecule-Substrate Interactions by Single Molecule Force Spectroscopy and Brownian Dynamics Simulations

    NASA Astrophysics Data System (ADS)

    Cook, Sara Iliafar

    Hybrids of biomolecules and nanomaterials have been identified as promising candidates in the development of novel therapeutics and electronic devices. Single stranded DNA (ssDNA)-bound Single-walled carbon nanotubes (SWCNTs) are of particular interest as they may be the key to solving the challenges that face the carbon nanotube separation technology and because of their potential application in bio-nanomedicine. The ability of ssDNA to form a stable hybrid with CNTs has been attributed to the structure and amphiphilic nature of this macromolecule, enabling the dispersion, sorting and patterned placement of nanotubes. Considering the significant role of ssDNA-CNTs in future technologies and the potential toxicity of such nanomaterials in biological systems, it is essential to gain a quantitative and fundamental understanding on the interactions that allow, weaken or prevent the formation of these hybrids. In this dissertation, we use both experimental and theoretical methods to systematically investigate the major characteristics of these interactions. The free energy of binding of ssDNA homopolymers to solvated carbon nanotubes is one of the key characteristics that determine the stability of such dispersions. We used single molecule force spectroscopy (SMFS), first on graphite and next on single walled carbon nanotubes, to probe and directly quantify the binding strength of ssDNA homopolymer oligomers to these substrates. The force resisting removal of DNA molecules from these surfaces shows characteristic steady-state force plateaus which were distinguishable for each DNA sequence. The free energy of binding per nucleotide for these oligomers on graphite were ranked as T >= A > G >= C (11.3 +/- 0.8 kT, 9.9 +/- 0.4 kT, 8.3 +/- 0.2 kT, and 7.5 +/- 0.8 kT, respectively). On SWCNTs, these interactions decreased in the following order: A > G > T > C, and their magnitude was much larger than on graphite (38.1 +/- 0.2; 33.9 +/- 0.1; 23.3 +/- 0.1; 17.1 +/- 0.1 k

  19. Single molecule force spectroscopy study of γ-polyglutamic acid by using atomic force microscopy

    NASA Astrophysics Data System (ADS)

    Ahamed, Parbhej; Kawakami, Masaru; Furukawa, Hidemitsu

    2016-04-01

    γ-PGA is a versatile multi-functional biopolymer possessing much useful biological functionality. Each biological function is associated with specific properties related to various forms of γ-PGA. However, to date, knowledge about the molecular mechanism to control specific structures of γ-PGA under a certain environmental conditions are unknown. In this study, we used single molecule force spectroscopy (SMFS) method to characterize the structure of γ-PGA in two environmental conditions. A water environment or 0.5 M MgCl2 salt solution was used to observe the effect of metal ion on γ-PGA structure at room temperature. The obtained results revealed that γ-PGA exists branched and/or cross-linked structure of γ-PGA in water or MgCl2 salt solution.

  20. Confocal Raman-AFM, A New Tool for Materials Research

    NASA Astrophysics Data System (ADS)

    Schmidt, Ute

    2005-03-01

    Characterization of heterogeneous systems, e.g. polymers, on the nanometer scale continues to grow in importance and to impact key applications in the field of materials science, nanotechnology and catalysis. The development of advanced polymeric materials for such applications requires detailed information about the physical and chemical properties of these materials on the nanometer scale. However, some details about the phase-separation process in polymers are difficult to study with conventional characterization techniques due to the inability of these methods to chemically differentiate materials with good spatial resolution, without damage, staining or preferential solvent washing. The CR-AFM is a breakthrough in microscopy. It combines three measuring techniques in one instrument: a high resolution confocal optical microscope, an extremely sensitive Raman spectroscopy system, and an Atomic Force Microscope. Using this instrument, the high spatial and topographical resolution obtained with an AFM can be directly linked to the chemical information gained by Confocal Raman spectroscopy. To demonstrate the capabilities of this unique combination of measuring techniques, polymer blend films, spin coated on glass substrates, have been characterized. AFM measurements reveal the structural and mechanical properties of the films, whereas Raman spectral images show the chemical composition of the blends.

  1. Versatile method for AFM-tip functionalization with biomolecules: fishing a ligand by means of an in situ click reaction

    NASA Astrophysics Data System (ADS)

    Kumar, Rakesh; Ramakrishna, Shivaprakash N.; Naik, Vikrant V.; Chu, Zonglin; Drew, Michael E.; Spencer, Nicholas D.; Yamakoshi, Yoko

    2015-04-01

    A facile and universal method for the functionalization of an AFM tip has been developed for chemical force spectroscopy (CFS) studies of intermolecular interactions of biomolecules. A click reaction between tripod-acetylene and an azide-linker-ligand molecule was successfully carried out on the AFM tip surface and used for the CFS study of ligand-receptor interactions.A facile and universal method for the functionalization of an AFM tip has been developed for chemical force spectroscopy (CFS) studies of intermolecular interactions of biomolecules. A click reaction between tripod-acetylene and an azide-linker-ligand molecule was successfully carried out on the AFM tip surface and used for the CFS study of ligand-receptor interactions. Electronic supplementary information (ESI) available: Experimental details with synthesis and characterization of compounds. Procedures for modifications of Au surfaces and AFM tips. AFM images and full PM-IRRAS spectra of modified surfaces. Detailed procedure for QCM measurement. A table showing ligand-receptor interaction probability. NMR, IR and MS charts. See DOI: 10.1039/c5nr01495f

  2. Understanding the plasmonics of nanostructured atomic force microscopy tips

    NASA Astrophysics Data System (ADS)

    Sanders, A.; Bowman, R. W.; Zhang, L.; Turek, V.; Sigle, D. O.; Lombardi, A.; Weller, L.; Baumberg, J. J.

    2016-10-01

    Structured metallic tips are increasingly important for optical spectroscopies such as tip-enhanced Raman spectroscopy, with plasmonic resonances frequently cited as a mechanism for electric field enhancement. We probe the local optical response of sharp and spherical-tipped atomic force microscopy (AFM) tips using a scanning hyperspectral imaging technique to identify the plasmonic behaviour. Localised surface plasmon resonances which radiatively couple with far-field light are found only for spherical AFM tips, with little response for sharp AFM tips, in agreement with numerical simulations of the near-field response. The precise tip geometry is thus crucial for plasmon-enhanced spectroscopies, and the typical sharp cones are not preferred.

  3. Age determination of blood spots in forensic medicine by force spectroscopy.

    PubMed

    Strasser, Stefan; Zink, Albert; Kada, Gerald; Hinterdorfer, Peter; Peschel, Oliver; Heckl, Wolfgang M; Nerlich, Andreas G; Thalhammer, Stefan

    2007-07-20

    We present a new tool for the estimation of the age of bloodstains, which could probably be used during forensic casework. For this, we used atomic force microscopy (AFM) for high-resolution imaging of erythrocytes in a blood sample and the detection of elasticity changes on a nanometer scale. For the analytic procedure we applied a fresh blood spot on a glass slide and started the AFM detection after drying of the blood drop. In a first step, an overview image was generated showing the presence of several red blood cells, which could easily be detected due to their typical "doughnut-like" appearance. The consecutively morphological investigations in a timeframe of 4 weeks could not show any alterations. Secondly, AFM was used to test the elasticity by recording force-distance curves. The measurements were performed immediately after drying, 1.5 h, 30 h and 31 days. The conditions were kept constant at room temperature (20 degrees C) and a humidity of 30%. The obtained elasticity parameters were plotted against a timeline and repeated several times. The elasticity pattern showed a decrease over time, which are most probably influenced by the alteration of the blood spot during the drying and coagulation process. The preliminary data demonstrates the capacity of this method to use it for development of calibration curves, which can be used for estimation of bloodstain ages during forensic investigations.

  4. Nanoscale fabrication of a peptide layer using an AFM probe

    NASA Astrophysics Data System (ADS)

    Nakamura, Chikashi; Miyamoto, Chie; Obataya, Ikuo; Nakamura, Noriyuki; Miyake, Jun

    2004-12-01

    Scanning probe microscopy has been applied in many studies to manipulate atoms or molecules. In particular, force spectroscopy using an atomic force microscope (AFM) is a powerful tool to elucidate intermolecular or intramolecular interactions and provide mechanical information. If enzymes could retain their activity when immobilized on probes, not only could enzyme-substrate interactions be investigated but also the probes could be used for precise biomolecular manipulation at the nano-scale. In our study, a method based on "Enzymatic Nanolithography" was successfully performed in a buffered solution using Staphylococcal serine V8 protease and AFM. To estimate the fabricating activity of the protease immobilized on the AFM tip to peptides immobilized on a substrate, we designed and synthesized peptides that showed enzymatic action specific to the protease. When the protease digested the reporter peptide a quencher residue was released from the main flame of the peptide and resulted in fluorescence. In the designed 9 mer peptides, TAMRA functioned as a good quencher for FAM. After contact of the protease-immobilized tip to the reporter peptide layer, a fluorescent area was observed by microscopic imaging.

  5. pH-dependent deformations of the energy landscape of avidin-like proteins investigated by single molecule force spectroscopy.

    PubMed

    Köhler, Melanie; Karner, Andreas; Leitner, Michael; Hytönen, Vesa P; Kulomaa, Markku; Hinterdorfer, Peter; Ebner, Andreas

    2014-08-18

    Avidin and avidin-like proteins are widely used in numerous techniques since the avidin-biotin interaction is known to be very robust and reliable. Within this study, we investigated this bond at the molecular level under harsh conditions ranging from very low to very high pH values. We compared avidin with streptavidin and a recently developed avidin-based mutant, chimeric avidin. To gain insights of the energy landscape of these interactions we used a single molecule approach and performed the Single Molecule Force Spectroscopy atomic force microscopy technique. There, the ligand (biotin) is covalently coupled to a sharp AFM tip via a distensible hetero-bi-functional crosslinker, whereas the receptor of interest is immobilized on the probe surface. Receptor-ligand complexes are formed and ruptured by repeatedly approaching and withdrawing the tip from the surface. Varying both pulling velocity and pH value, we could determine changes of the energy landscape of the complexes. Our results clearly demonstrate that avidin, streptavidin and chimeric avidin are stable over a wide pH range although we could identify differences at the outer pH range. Taking this into account, they can be used in a broad range of applications, like surface sensors at extreme pH values.

  6. AFM cantilever vibration detection with a transmitted electron beam

    NASA Astrophysics Data System (ADS)

    Woehl, Taylor; Wagner, Ryan; Keller, Robert; Killgore, Jason

    Cantilever oscillations for dynamic atomic force microscopy (AFM) are conventionally measured with an optical lever system. The speed of AFM cantilevers can be increased by decreasing the size of the cantilever; however, the fastest AFM cantilevers are currently nearing the smallest size that can be detected with the current optical lever approach. Here we demonstrate an electron detection scheme in an SEM for detecting AFM cantilever oscillations. An oscillating AFM tip is positioned perpendicular to the propagation direction of a stationary ~ 1 nm diameter electron probe, and the oscillatory change in electron scattering resulting from the changing thickness of the electron irradiated area of the AFM tip is detected with a transmitted electron detector positioned below the AFM tip. We perform frequency sweep and ring-down experiments to determine the first resonant frequency and Q factor of an AFM cantilever.

  7. Study of Thin Oxide Films with NC-AFM: Atomically Resolved Imaging and Beyond

    NASA Astrophysics Data System (ADS)

    Heyde, M.; Simon, G. H.; König, T.

    Results presented in the following show structural analysis of metaloxide surfaces and the extraction of physical quantities from the force field above such a surface by noncontact atomic force microscopy (NC-AFM). The measurements have been performed with our dual mode NC-AFM/STM in ultrahigh vacuum at 5 K. The introduction will be followed by a description of the experimental setup, including the ultrahigh vacuum cryogenic environment and our tuning fork tunneling current and force sensor. The sensor parameters affecting the measurements are given together with an amplitude characterization method. In the next section, a structure determination of ultrathin Alumina/NiAl(110) is shown. Atomic resolution could be achieved throughout both reflection domain unit cells. NC-AFM reveals details of morphological features, interconnections to substrate-film interactions, and comparability to theory also with respect to topographic height. In the last section, we present measurements beyond imaging, namely spectroscopy data taken on thin MgO films grown on Ag(001). Force-distance measurements based on atomically resolved NC-AFM images of these films have been taken. Inequivalent sites could be resolved and their effect on nucleation and adsorption processes is debated. Furthermore, work function shift measurements on different MgO film thicknesses grown on Ag(001) are studied and the impact of this shift on the catalytic properties of adsorbed metal species is discussed.

  8. Microfluidics, Chromatography, and Atomic-Force Microscopy

    NASA Technical Reports Server (NTRS)

    Anderson, Mark

    2008-01-01

    A Raman-and-atomic-force microscope (RAFM) has been shown to be capable of performing several liquid-transfer and sensory functions essential for the operation of a microfluidic laboratory on a chip that would be used to perform rapid, sensitive chromatographic and spectro-chemical analyses of unprecedentedly small quantities of liquids. The most novel aspect of this development lies in the exploitation of capillary and shear effects at the atomic-force-microscope (AFM) tip to produce shear-driven flow of liquids along open microchannels of a microfluidic device. The RAFM can also be used to perform such functions as imaging liquids in microchannels; removing liquid samples from channels for very sensitive, tip-localized spectrochemical analyses; measuring a quantity of liquid adhering to the tip; and dip-pen deposition from a chromatographic device. A commercial Raman-spectroscopy system and a commercial AFM were integrated to make the RAFM so as to be able to perform simultaneous topographical AFM imaging and surface-enhanced Raman spectroscopy (SERS) at the AFM tip. The Raman-spectroscopy system includes a Raman microprobe attached to an optical microscope, the translation stage of which is modified to accommodate the AFM head. The Raman laser excitation beam, which is aimed at the AFM tip, has a wavelength of 785 nm and a diameter of about 5 m, and its power is adjustable up to 10 mW. The AFM is coated with gold to enable tip-localized SERS.

  9. Condensed-phase thermal decomposition of TATB investigated by atomic force microscopy (AFM) and simultaneous thermogravimetric modulated beam mass spectrometry (STMBMS)

    SciTech Connect

    Land, T.A.; Siekhaus, W.J.; Foltz, M.F.; Behrens, R. Jr.

    1993-05-01

    A combination of techniques has been used to investigate the condensed-phase thermal decomposition of TATB. STMBMS has been used to identify the thermal decomposition products and their temporal correlation`s. These experiments have shown that the condensed-phase decomposition proceeds through several autocatalytic pathways. Both low and high molecular weight decomposition products have been identified. Mono-, di- and tri-furazans products have been identified and, their temporal behaviors are consistent with a stepwise loss of water. AFM has been used to correlate the decomposition chemistry with morphological changes occurring as a function of heating. Patches of small 25-140 nm round holes were observed throughout the lattice of TATB crystals that were heated briefly to 300C. It is likely that these holes show where decomposition reactions have started. Evidence of decomposition products have been seen in TATB that has been held at 250C for one hour.

  10. Nano Mechanical Machining Using AFM Probe

    NASA Astrophysics Data System (ADS)

    Mostofa, Md. Golam

    Complex miniaturized components with high form accuracy will play key roles in the future development of many products, as they provide portability, disposability, lower material consumption in production, low power consumption during operation, lower sample requirements for testing, and higher heat transfer due to their very high surface-to-volume ratio. Given the high market demand for such micro and nano featured components, different manufacturing methods have been developed for their fabrication. Some of the common technologies in micro/nano fabrication are photolithography, electron beam lithography, X-ray lithography and other semiconductor processing techniques. Although these methods are capable of fabricating micro/nano structures with a resolution of less than a few nanometers, some of the shortcomings associated with these methods, such as high production costs for customized products, limited material choices, necessitate the development of other fabricating techniques. Micro/nano mechanical machining, such an atomic force microscope (AFM) probe based nano fabrication, has, therefore, been used to overcome some the major restrictions of the traditional processes. This technique removes material from the workpiece by engaging micro/nano size cutting tool (i.e. AFM probe) and is applicable on a wider range of materials compared to the photolithographic process. In spite of the unique benefits of nano mechanical machining, there are also some challenges with this technique, since the scale is reduced, such as size effects, burr formations, chip adhesions, fragility of tools and tool wear. Moreover, AFM based machining does not have any rotational movement, which makes fabrication of 3D features more difficult. Thus, vibration-assisted machining is introduced into AFM probe based nano mechanical machining to overcome the limitations associated with the conventional AFM probe based scratching method. Vibration-assisted machining reduced the cutting forces

  11. APOBEC3G Interacts with ssDNA by Two Modes: AFM Studies

    NASA Astrophysics Data System (ADS)

    Shlyakhtenko, Luda S.; Dutta, Samrat; Banga, Jaspreet; Li, Ming; Harris, Reuben S.; Lyubchenko, Yuri L.

    2015-10-01

    APOBEC3G (A3G) protein has antiviral activity against HIV and other pathogenic retroviruses. A3G has two domains: a catalytic C-terminal domain (CTD) that deaminates cytidine, and a N-terminal domain (NTD) that binds to ssDNA. Although abundant information exists about the biological activities of A3G protein, the interplay between sequence specific deaminase activity and A3G binding to ssDNA remains controversial. We used the topographic imaging and force spectroscopy modalities of Atomic Force Spectroscopy (AFM) to characterize the interaction of A3G protein with deaminase specific and nonspecific ssDNA substrates. AFM imaging demonstrated that A3G has elevated affinity for deaminase specific ssDNA than for nonspecific ssDNA. AFM force spectroscopy revealed two distinct binding modes by which A3G interacts with ssDNA. One mode requires sequence specificity, as demonstrated by stronger and more stable complexes with deaminase specific ssDNA than with nonspecific ssDNA. Overall these observations enforce prior studies suggesting that both domains of A3G contribute to the sequence specific binding of ssDNA.

  12. Multiplexed single-molecule force spectroscopy using a centrifuge

    NASA Astrophysics Data System (ADS)

    Yang, Darren; Ward, Andrew; Halvorsen, Ken; Wong, Wesley P.

    2016-03-01

    We present a miniature centrifuge force microscope (CFM) that repurposes a benchtop centrifuge for high-throughput single-molecule experiments with high-resolution particle tracking, a large force range, temperature control and simple push-button operation. Incorporating DNA nanoswitches to enable repeated interrogation by force of single molecular pairs, we demonstrate increased throughput, reliability and the ability to characterize population heterogeneity. We perform spatiotemporally multiplexed experiments to collect 1,863 bond rupture statistics from 538 traceable molecular pairs in a single experiment, and show that 2 populations of DNA zippers can be distinguished using per-molecule statistics to reduce noise.

  13. Multiplexed single-molecule force spectroscopy using a centrifuge.

    PubMed

    Yang, Darren; Ward, Andrew; Halvorsen, Ken; Wong, Wesley P

    2016-01-01

    We present a miniature centrifuge force microscope (CFM) that repurposes a benchtop centrifuge for high-throughput single-molecule experiments with high-resolution particle tracking, a large force range, temperature control and simple push-button operation. Incorporating DNA nanoswitches to enable repeated interrogation by force of single molecular pairs, we demonstrate increased throughput, reliability and the ability to characterize population heterogeneity. We perform spatiotemporally multiplexed experiments to collect 1,863 bond rupture statistics from 538 traceable molecular pairs in a single experiment, and show that 2 populations of DNA zippers can be distinguished using per-molecule statistics to reduce noise. PMID:26984516

  14. Multiplexed single-molecule force spectroscopy using a centrifuge

    PubMed Central

    Yang, Darren; Ward, Andrew; Halvorsen, Ken; Wong, Wesley P.

    2016-01-01

    We present a miniature centrifuge force microscope (CFM) that repurposes a benchtop centrifuge for high-throughput single-molecule experiments with high-resolution particle tracking, a large force range, temperature control and simple push-button operation. Incorporating DNA nanoswitches to enable repeated interrogation by force of single molecular pairs, we demonstrate increased throughput, reliability and the ability to characterize population heterogeneity. We perform spatiotemporally multiplexed experiments to collect 1,863 bond rupture statistics from 538 traceable molecular pairs in a single experiment, and show that 2 populations of DNA zippers can be distinguished using per-molecule statistics to reduce noise. PMID:26984516

  15. AFM characterization of nanobubble formation and slip condition in oxygenated and electrokinetically altered fluids.

    PubMed

    Bhushan, Bharat; Pan, Yunlu; Daniels, Stephanie

    2013-02-15

    Nanobubbles are gas-filled features that spontaneously form at the interface of hydrophobic surfaces and aqueous solutions. In this study, an atomic force microscope (AFM) was used to characterize the morphology of nanobubbles formed on hydrophobic polystyrene (PS) and octadecyltrichlorosilane (OTS) films immersed in DI water, saline, saline with oxygen and an electrokinetically altered saline solution produced with Taylor-Couette-Poiseuille flow under elevated oxygen pressure. AFM force spectroscopy was used to evaluate hydrodynamic and electrostatic forces and boundary slip condition in various fluids. The effect of solution, electric field and surface charge on shape, size and density of nanobubbles as well as slip length was quantified and the results and underlying mechanisms are presented in this paper.

  16. On the molecular interaction between albumin and ibuprofen: An AFM and QCM-D study.

    PubMed

    Eleta-Lopez, Aitziber; Etxebarria, Juan; Reichardt, Niels-Christian; Georgieva, Radostina; Bäumler, Hans; Toca-Herrera, José L

    2015-10-01

    The adsorption of proteins on surfaces often results in a change of their structural behavior and consequently, a loss of bioactivity. One experimental method to study interactions on a molecular level is single molecular force spectroscopy that permits to measure forces down to the pico-newton range. In this work, the binding force between human serum albumin (HSA), covalently immobilized on glutaraldehyde modified gold substrates, and ibuprofen sodium salt was studied by means of single molecular force spectroscopy. First of all, a protocol was established to functionalize atomic force microscopy (AFM) tips with ibuprofen. The immobilization protocol was additionally tested by quartz crystal microbalance with dissipation (QCM-D) and contact angle measurements. AFM was used to characterize the adsorption of HSA on gold substrates, which lead to a packed monolayer of thickness slightly lower than the reported value in solution. Finally, single molecule spectroscopy results were used to characterize the binding force between albumin and ibuprofen and calculate the distance of the transition state (0.6 nm) and the dissociation rate constant (0.055 s(-1)). The results might indicate that part of the adsorbed protein still preserves its functionality upon adsorption.

  17. On the molecular interaction between albumin and ibuprofen: An AFM and QCM-D study.

    PubMed

    Eleta-Lopez, Aitziber; Etxebarria, Juan; Reichardt, Niels-Christian; Georgieva, Radostina; Bäumler, Hans; Toca-Herrera, José L

    2015-10-01

    The adsorption of proteins on surfaces often results in a change of their structural behavior and consequently, a loss of bioactivity. One experimental method to study interactions on a molecular level is single molecular force spectroscopy that permits to measure forces down to the pico-newton range. In this work, the binding force between human serum albumin (HSA), covalently immobilized on glutaraldehyde modified gold substrates, and ibuprofen sodium salt was studied by means of single molecular force spectroscopy. First of all, a protocol was established to functionalize atomic force microscopy (AFM) tips with ibuprofen. The immobilization protocol was additionally tested by quartz crystal microbalance with dissipation (QCM-D) and contact angle measurements. AFM was used to characterize the adsorption of HSA on gold substrates, which lead to a packed monolayer of thickness slightly lower than the reported value in solution. Finally, single molecule spectroscopy results were used to characterize the binding force between albumin and ibuprofen and calculate the distance of the transition state (0.6 nm) and the dissociation rate constant (0.055 s(-1)). The results might indicate that part of the adsorbed protein still preserves its functionality upon adsorption. PMID:26218522

  18. SU-8 hollow cantilevers for AFM cell adhesion studies

    NASA Astrophysics Data System (ADS)

    Martinez, Vincent; Behr, Pascal; Drechsler, Ute; Polesel-Maris, Jérôme; Potthoff, Eva; Vörös, Janos; Zambelli, Tomaso

    2016-05-01

    A novel fabrication method was established to produce flexible, transparent, and robust tipless hollow atomic force microscopy (AFM) cantilevers made entirely from SU-8. Channels of 3 μm thickness and several millimeters length were integrated into 12 μm thick and 40 μm wide cantilevers. Connected to a pressure controller, the devices showed high sealing performance with no leakage up to 6 bars. Changing the cantilever lengths from 100 μm to 500 μm among the same wafer allowed the targeting of various spring constants ranging from 0.5 to 80 N m-1 within a single fabrication run. These hollow polymeric AFM cantilevers were operated in the optical beam deflection configuration. To demonstrate the performance of the device, single-cell force spectroscopy experiments were performed with a single probe detaching in a serial protocol more than 100 Saccharomyces cerevisiae yeast cells from plain glass and glass coated with polydopamine while measuring adhesion forces in the sub-nanoNewton range. SU-8 now offers a new alternative to conventional silicon-based hollow cantilevers with more flexibility in terms of complex geometric design and surface chemistry modification.

  19. SU-8 hollow cantilevers for AFM cell adhesion studies

    NASA Astrophysics Data System (ADS)

    Martinez, Vincent; Behr, Pascal; Drechsler, Ute; Polesel-Maris, Jérôme; Potthoff, Eva; Vörös, Janos; Zambelli, Tomaso

    2016-05-01

    A novel fabrication method was established to produce flexible, transparent, and robust tipless hollow atomic force microscopy (AFM) cantilevers made entirely from SU-8. Channels of 3 μm thickness and several millimeters length were integrated into 12 μm thick and 40 μm wide cantilevers. Connected to a pressure controller, the devices showed high sealing performance with no leakage up to 6 bars. Changing the cantilever lengths from 100 μm to 500 μm among the same wafer allowed the targeting of various spring constants ranging from 0.5 to 80 N m‑1 within a single fabrication run. These hollow polymeric AFM cantilevers were operated in the optical beam deflection configuration. To demonstrate the performance of the device, single-cell force spectroscopy experiments were performed with a single probe detaching in a serial protocol more than 100 Saccharomyces cerevisiae yeast cells from plain glass and glass coated with polydopamine while measuring adhesion forces in the sub-nanoNewton range. SU-8 now offers a new alternative to conventional silicon-based hollow cantilevers with more flexibility in terms of complex geometric design and surface chemistry modification.

  20. Electron work functions of ferrite and austenite phases in a duplex stainless steel and their adhesive forces with AFM silicon probe

    PubMed Central

    Guo, Liqiu; Hua, Guomin; Yang, Binjie; Lu, Hao; Qiao, Lijie; Yan, Xianguo; Li, Dongyang

    2016-01-01

    Local electron work function, adhesive force, modulus and deformation of ferrite and austenite phases in a duplex stainless steel were analyzed by scanning force microscopy. It is demonstrated that the austenite has a higher electron work function than the ferrite, corresponding to higher modulus, smaller deformation and larger adhesive force. Relevant first-principles calculations were conducted to elucidate the mechanism behind. It is demonstrated that the difference in the properties between austenite and ferrite is intrinsically related to their electron work functions. PMID:26868719

  1. The NTA-His6 bond is strong enough for AFM single-molecular recognition studies.

    PubMed

    Verbelen, Claire; Gruber, Hermann J; Dufrêne, Yves F

    2007-01-01

    There is a need in current atomic force microscopy (AFM) molecular recognition studies for generic methods for the stable, functional attachment of proteins on tips and solid supports. In the last few years, the site-directed nitrilotriacetic acid (NTA)-polyhistidine (Hisn) system has been increasingly used towards this goal. Yet, a crucial question in this context is whether the NTA-Hisn bond is sufficiently strong for ensuring stable protein immobilization during force spectroscopy measurements. Here, we measured the forces between AFM tips modified with NTA-terminated alkanethiols and solid supports functionalized with His6-Gly-Cys peptides in the presence of Ni2+. The force histogram obtained at a loading rate of 6600 pN s(-1) showed three maxima at rupture forces of 153 +/- 57 pN, 316 +/- 50 pN and 468 +/- 44 pN, that we attribute primarily to monovalent and multivalent interactions between a single His6 moiety and one, two and three NTA groups, respectively. The measured forces are well above the 50-100 pN unbinding forces typically observed by AFM for receptor-ligand pairs. The plot of adhesion force versus log (loading rate) revealed a linear regime, from which we deduced a kinetic off-rate constant of dissociation, k(off) approximately 0.07 s(-1). This value is in the range of that estimated for the multivalent interaction involving two NTA, using fluorescence measurements, and may account for an increased binding stability of the NTA-His6 bond. We conclude that the NTA-His6 system is a powerful, well-suited platform for the stable, oriented immobilization of proteins in AFM single-molecule studies. PMID:17712775

  2. Unbinding forces and energies between a siRNA molecule and a dendrimer measured by force spectroscopy.

    PubMed

    Dumitru, Andra C; Herruzo, Elena T; Rausell, Estrella; Ceña, Valentin; Garcia, Ricardo

    2015-12-21

    We have measured the intermolecular forces between small interference RNA (siRNA) and polyamidoamine dendrimers at the single molecular level. A single molecule force spectroscopy approach has been developed to measure the unbinding forces and energies between a siRNA molecule and polyamidoamine dendrimers deposited on a mica surface in a buffer solution. We report three types of unbinding events which are characterized by forces and free unbinding energies, respectively, of 28 pN, 0.709 eV; 38 pN, 0.722 eV; and 50 pN, 0.724 eV. These events reflect different possible electrostatic interactions between the positive charges of one or two dendrimers and the negatively charged phosphate groups of a single siRNA. We have evidence of a high binding affinity of siRNA towards polyamidoamine dendrimers that leads to a 45% probability of measuring specific unbinding events.

  3. Tuning the Music: Acoustic Force Spectroscopy (AFS) 2.0.

    PubMed

    Kamsma, Douwe; Creyghton, Ramon; Sitters, Gerrit; Wuite, Gijs J L; Peterman, Erwin J G

    2016-08-01

    AFS is a recently introduced high-throughput single-molecule technique that allows studying structural and mechanochemical properties of many biomolecules in parallel. To further improve the method, we developed a modelling tool to optimize the layer thicknesses, and a calibration method to experimentally validate the modelled force profiles. After optimization, we are able to apply 350pN on 4.5μm polystyrene beads, without the use of an amplifier, at the coverslip side of the AFS chip. Furthermore, we present the use of a transparent piezo to generate the acoustic force and we show that AFS can be combined with high-NA oil or water-immersion objectives. With this set of developments AFS will be applicable to a broad range of single-molecule experiments. PMID:27163865

  4. Atom-specific forces and defect identification on surface-oxidized Cu(100) with combined 3D-AFM and STM measurements

    NASA Astrophysics Data System (ADS)

    Baykara, Mehmet Z.; Todorović, Milica; Mönig, Harry; Schwendemann, Todd C.; Ünverdi, Özhan; Rodrigo, Lucia; Altman, Eric I.; Pérez, Rubén; Schwarz, Udo D.

    2013-04-01

    The influence of defects on the local structural, electronic, and chemical properties of a surface oxide on Cu(100) were investigated using atomic resolution three-dimensional force mapping combined with tunneling current measurements and ab initio density functional theory. Results reveal that the maximum attractive force between tip and sample occurs above the oxygen atoms; theory indicates that the tip, in this case, terminates in a Cu atom. Meanwhile, simultaneously acquired tunneling current images emphasize the positions of Cu atoms, thereby, providing species-selective contrast in the two complementary data channels. One immediate outcome is that defects due to the displacement of surface copper are exposed in the current maps, even though force maps only reflect a well-ordered oxygen sublattice. The exact nature of the defects is confirmed by the simulations, which also reveal that the arrangement of the oxygen atoms is not disrupted by the copper displacement. In addition, the experimental force maps uncover a position-dependent modulation of the attractive forces between the surface oxygen and the copper-terminated tips, which is found to reflect the surface's inhomogeneous chemical and structural environment. As a consequence, the demonstrated method has the potential to directly probe how defects affect surface chemical interactions.

  5. Mapping the local dielectric response at the nanoscale by means of plasmonic force spectroscopy.

    PubMed

    De Angelis, Francesco; Zaccaria, Remo Proietti; Di Fabrizio, Enzo

    2012-12-31

    At the present, the local optical properties of nanostructured materials are difficult to be measured by available instrumentation. We investigated the capability of plasmonic force spectroscopy of measuring the optical response at the nanoscale. The proposed technique is based on force measurements performed by combining Atomic Force Microscopy, or optical tweezers, and adiabatic compression of surface plasmon polaritons. We show that the optical forces, caused by the plasmonic field, depend on the local response of the substrates and, in principle, allow probing both the real and the imaginary part of the local permittivity with a spatial resolution of few nanometers.

  6. Molecular force spectroscopy of homophilic nectin-1 interactions

    SciTech Connect

    Vedula, Sri Ram Krishna; Lim, T.S.; Hui Shi; Kausalya, P. Jaya; Lane, E. Birgitte; Rajagopal, Gunaretnam; Hunziker, Walter; Lim, C.T.

    2007-11-03

    Nectins are Ca{sup 2+} independent cell adhesion molecules localizing at the cadherin based adherens junctions. In this study, we have used atomic force microscopy to study interaction of a chimera of extra cellular fragment of nectin-1 and Fc of human IgG (nef-1) with wild type L-fibroblasts that express endogenous nectin-1 to elucidate the biophysical characteristics of homophilic nectin-1 trans-interactions at the level of single molecule. Bond strength distribution revealed three distinct bound states (or configurations) of trans-interactions between paired nectins, where each bound state has a unique unstressed off-rate and reactive compliance. Kinetic analysis of force-dependent off-rate of the bound state involving trans-interacting V-V domains between paired nectin-1 (unstressed off-rate {approx}1.465 {+-} 0.779 s{sup -1}, reactive compliance {approx}0.143 {+-} 0.072 nm) was found to be closest to E-cadherin, indicating that V-V domain trans-interactions are probably necessary to initiate and promote adhesions of E-cadherin at adherens junctions (AJs)

  7. Measurement of the interaction forces at various pH levels by using AFM for the interpretation of DNA adsorption on silanized surfaces

    NASA Astrophysics Data System (ADS)

    Han, Seung Pil; Suga, Kosaku; Fujihara, Masamichi; Park, Byung-Eun

    2014-09-01

    Various surfaces have been used for deoxyribonucleic acid (DNA) immobilization, one example being a silanized surface. This is useful for determining DNA lengths and, thus, locating specific gene sequences in DNA by using fluorescence microscopy and scanning probe microscopy. In this study, we deposited DNA by using the molecular combing method and, we used fluorescence microscopy to study how the chain lengths of n-alkylsilanes affected the surface density of DNA deposited on the silanized surfaces in a tris-ethylenediaminetetraacetic acid (TE) buffer. The forces between a cleaned silicon-nitride (Si3N4) tip and each substrate surface in aqueous buffers at various pH levels (1.0 ~ 9.0) were also studied by using atomic force microscopy to measure the force-distance curves. We explain why the density of lambda bacteriophage DNA (λ-DNA) deposited by using the molecular combing method at pH 8 was lower on the silanized surface with the shorter alkyl chain than it was on the silanized surface with the longer alkyl chain in terms of the electrical double layer (EDL) and the adhesive force.

  8. Direct AFM force measurements between air bubbles in aqueous polydisperse sodium poly(styrene sulfonate) solutions: effect of collision speed, polyelectrolyte concentration and molar mass.

    PubMed

    Browne, Christine; Tabor, Rico F; Grieser, Franz; Dagastine, Raymond R

    2015-07-01

    Interactions between colliding air bubbles in aqueous solutions of polydisperse sodium poly(styrene sulfonate) (NaPSS) using direct force measurements were studied. The forces measured with deformable interfaces were shown to be more sensitive to the presence of the polyelectrolytes when compared to similar measurements using rigid interfaces. The experimental factors that were examined were NaPSS concentration, bubble collision velocity and polyelectrolyte molar mass. These measurements were then compared with an analytical model based on polyelectrolyte scaling theory in order to explain the effects of concentration and bubble deformation on the interaction between bubbles. Typically structural forces from the presence of monodisperse polyelectrolyte between interacting surfaces may be expected, however, it was found that the polydispersity in molar mass resulted in the structural forces to be smoothed and only a depletion interaction was able to be measured between interacting bubbles. It was found that an increase in number density of NaPSS molecules resulted in an increase in the magnitude of the depletion interaction. Conversely this interaction was overwhelmed by an increase in the fluid flow in the system at higher bubble collision velocities. Polymer molar mass dispersity plays a significant role in the interactions present between the bubbles and has implications that also affect the polyelectrolyte overlap concentration of the solution. Further understanding of these implications can be expected to play a role in the improvement in operations in such fields as water treatment and mineral processing where polyelectrolytes are used extensively.

  9. Quantitative Atomic-Resolution Surface Force Field Spectroscopy in Three Dimensions: A How-To Guide for Collecting Meaningful Data

    NASA Astrophysics Data System (ADS)

    Baykara, Mehmet Z.; Dagdeviren, Omur E.; Schwendemann, Todd C.; MöNig, Harry; Altman, Eric I.; Schwarz, Udo D.

    2013-03-01

    Three-dimensional atomic force microscopy (3D-AFM) is being increasingly used to measure the chemical interactions between an atomically sharp probe tip and surfaces of interest in terms of atomic-scale forces and energies in three dimensions. Since the results provided by 3D-AFM may be affected by piezo nonlinearities, thermal and electronic drift, tip asymmetries, and elastic deformation of the tip's apex, these effects need to be considered during data interpretation. In this talk, we analyze the impact of these effects on the data, compare different methods to record atomic-resolution surface force fields, and determine the approaches that suffer the least from associated artifacts. We conclude that efforts to reduce unwanted influence of tip properties on recorded data are indispensable to extract detailed information about atomic-scale properties of the surface.

  10. Effect of molecular weight on the exponential growth and morphology of hyaluronan/chitosan multilayers: a surface plasmon resonance spectroscopy and atomic force microscopy investigation.

    PubMed

    Kujawa, Piotr; Moraille, Patricia; Sanchez, Jacqueline; Badia, Antonella; Winnik, Françoise M

    2005-06-29

    The layer-by-layer growth of multilayer assemblies of two polysaccharides, the polyanion hyaluronan (HA) and the polycation chitosan (CH), was investigated using atomic force microscopy (AFM) and surface plasmon resonance (SPR) spectroscopy, with primary emphasis on the effect of the polysaccharide molecular weights on the film thickness and surface morphology. The HA/CH multilayers exhibit an exponential increase of the optical film thickness with the number of deposited bilayers. We show that the multilayer thickness at a given stage depends on the size of both CH, the diffusing polyelectrolyte, and HA, the non-diffusing species. Assemblies (12 bilayers) of high molecular weight polysaccharides (HA, 360,000; CH, 160,000) were twice as thick (approximately 900 nm vs approximately 450 nm) as those obtained with low molecular weight polymers (HA, 30,000; CH, 31,000), as assessed by AFM scratch tests. The exponential growth rate is the same for the high and low molecular weight pairs; the larger film thicknesses observed by SPR and by AFM arising from an earlier onset of the steep exponential growth phase in the case of the high molecular weight pair. In all cases, isolated islets form during the deposition of the first CH layer onto the underlying HA. Upon further film growth, individual islets coalesce into larger vermiculate features. The transition from distinct islands to vermiculate structures depends on the molecular weights of the polysaccharides and the lower molecular weight construct presents larger worm-like surface domains than the high molecular weight pair.

  11. Surface electrical properties of stainless steel fibres: An AFM-based study

    NASA Astrophysics Data System (ADS)

    Yin, Jun; D'Haese, Cécile; Nysten, Bernard

    2015-03-01

    Atomic force microscopy (AFM) electrical modes were used to study the surface electrical properties of stainless steel fibres. The surface electrical conductivity was studied by current sensing AFM and I-V spectroscopy. Kelvin probe force microscopy was used to measure the surface contact potential. The oxide film, known as passivation layer, covering the fibre surface gives rise to the observation of an apparently semiconducting behaviour. The passivation layer generally exhibits a p-type semiconducting behaviour, which is attributed to the predominant formation of chromium oxide on the surface of the stainless steel fibres. At the nanoscale, different behaviours are observed from points to points, which may be attributed to local variations of the chemical composition and/or thickness of the passivation layer. I-V curves are well fitted with an electron tunnelling model, indicating that electron tunnelling may be the predominant mechanism for electron transport.

  12. Force spectroscopy of single multidomain biopolymers: A master equation approach

    NASA Astrophysics Data System (ADS)

    Braun, O.; Seifert, U.

    2005-09-01

    Experiments using atomic force microscopy for unfolding single multidomain biopolymers cover a broad range of time scales from equilibrium to non-equilibrium. A master equation approach allows to identify and treat coherently three dynamical regimes for increasing linear ramp velocity: i) an equilibrium regime, ii) a transient regime where refolding events still occur, and iii) a saw-tooth regime without any refolding events. For each regime, analytical approximations are derived and compared to numerically investigated examples. We analyze in the framework of this model also a periodic experimental protocol instead of a linear ramp. In this case, a major simplification arises if the dynamics can be restricted to an effectively two-dimensional subspace. For transitions with an intermediate meta-stable state, like Immunoglobulin27, a refined model allows to extract previously unknown molecular parameters related to this meta-stable state.

  13. Rheology of fluids measured by correlation force spectroscopy

    NASA Astrophysics Data System (ADS)

    Radiom, Milad; Robbins, Brian; Honig, Christopher D. F.; Walz, John Y.; Paul, Mark R.; Ducker, William A.

    2012-04-01

    We describe a method, correlation force spectrometry (CFS), which characterizes fluids through measurement of the correlations between the thermally stimulated vibrations of two closely spaced micrometer-scale cantilevers in fluid. We discuss a major application: measurement of the rheological properties of fluids at high frequency and high spatial resolution. Use of CFS as a rheometer is validated by comparison between experimental data and finite element modeling of the deterministic ring-down of cantilevers using the known viscosity of fluids. The data can also be accurately fitted using a harmonic oscillator model, which can be used for rapid rheometric measurements after calibration. The method is non-invasive, uses a very small amount of fluid, and has no actively moving parts. It can also be used to analyze the rheology of complex fluids. We use CFS to show that (non-Newtonian) aqueous polyethylene oxide solution can be modeled approximately by incorporating an elastic spring between the cantilevers.

  14. High spatial resolution surface imaging and analysis of fungal cells using SEM and AFM.

    PubMed

    Kaminskyj, Susan G W; Dahms, Tanya E S

    2008-06-01

    We review the use of scanning electron microscopy (SEM), atomic force microscopy (AFM) and force spectroscopy (FS) for probing the ultrastructure, chemistry, physical characteristics and motion of fungal cells. When first developed, SEM was used to image fixed/dehydrated/gold coated specimens, but here we describe more recent SEM developments as they apply to fungal cells. CryoSEM offers high resolution for frozen fungal samples, whereas environmental SEM allows the analysis of robust samples (e.g. spores) under ambient conditions. Dual beam SEM, the most recently developed, adds manipulation capabilities along with element detection. AFM has similar lateral and better depth resolution compared to SEM, and can image live cells including growing fungal hyphae. FS can analyze cell wall chemistry, elasticity and dynamic cell characteristics. The integration of AFM with optical microscopy will allow examination of individual molecules or cellular structures in the context of fungal cell architecture. SEM and AFM are complementary techniques that are clarifying our understanding of fungal biology. PMID:18068995

  15. Spatial spectrograms of vibrating atomic force microscopy cantilevers coupled to sample surfaces

    SciTech Connect

    Wagner, Ryan; Raman, Arvind; Proksch, Roger

    2013-12-23

    Many advanced dynamic Atomic Force Microscopy (AFM) techniques such as contact resonance, force modulation, piezoresponse force microscopy, electrochemical strain microscopy, and AFM infrared spectroscopy exploit the dynamic response of a cantilever in contact with a sample to extract local material properties. Achieving quantitative results in these techniques usually requires the assumption of a certain shape of cantilever vibration. We present a technique that allows in-situ measurements of the vibrational shape of AFM cantilevers coupled to surfaces. This technique opens up unique approaches to nanoscale material property mapping, which are not possible with single point measurements alone.

  16. Structural and Mechanical Mechanisms of Ocular Tissues Probed by AFM

    NASA Astrophysics Data System (ADS)

    Ziebarth, Noël M.; Rico, Felix; Moy, Vincent T.

    In recent years, the atomic force microscope (AFM) has become an important tool in ophthalmic research. It has gained popularity largely because AFM is not restricted by the diffraction limits of light microscopy and can be applied to resolve images with molecular resolution. AFM is a minimally invasive technique and can be used to visualize molecular structures under near-physiological conditions. In addition, the AFM can be employed as a force apparatus to characterize the viscoelastic properties of biomaterials on the micron level and at the level of individual proteins. In this article, we summarize recent AFM studies of ocular tissues, while highlighting the great potential of AFM technology in ophthalmic research. Previous research demonstrates the versatility of the AFM as high resolution imaging technique and as a sensitive force apparatus for probing the mechanical properties of ocular tissues. The structural and mechanical properties of ocular tissues are of major importance to the understanding of the optomechanical functions of the human eye. In addition, AFM has played an important role in the development and characterization of ocular biomaterials, such as contact lenses and intraocular lenses. Studying ocular tissues using Atomic Force Microscopy has enabled several advances in ophthalmic research.

  17. Probing the Higgs force with isotope shift spectroscopy

    NASA Astrophysics Data System (ADS)

    Ozeri, Roee; Delaunay, Cedric; Perez, Gilad; Soreq, Yotam

    2016-05-01

    The Higgs boson, the last missing piece of the Standard Model (SM) of elementary particles, was recently observed by experiments in the Large Hadron Collider (LHC). To check whether this is indeed the SM Higgs, its coupling to other elementary particles should be experimentally measured. Current limits placed by LHC experiments on the coupling of the Higgs to the main building block of matter; the electron and the up and down quarks; are orders of magnitude larger than the SM predictions. Here, we propose to use the measurement of isotope shifts in optical atomic clock transitions to probe the Higgs boson coupling to electrons and nuclei. We show that the Higgs force between nuclei and bound electrons induces measurable nonlinearities to the King relation between isotope shifts. With current state-of-the-art accuracy in frequency comparison, limits which compete with, or even surpass, the bounds provided by LHC experiments can be achieved. Improved knowledge of these couplings is an important test of the SM. Similarly, this measurement could lead to an improved sensitivity to the presence of new physics.

  18. Structural insight into iodide uptake by AFm phases.

    PubMed

    Aimoz, Laure; Wieland, Erich; Taviot-Guého, Christine; Dähn, Rainer; Vespa, Marika; Churakov, Sergey V

    2012-04-01

    The ability of cement phases carrying positively charged surfaces to retard the mobility of (129)I, present as iodide (I(-)) in groundwater, was investigated in the context of safe disposal of radioactive waste. (125)I sorption experiments on ettringite, hydrotalcite, chloride-, carbonate- and sulfate-containing AFm phases indicated that calcium-monosulfate (AFm-SO(4)) is the only phase that takes up trace levels of iodide. The structures of AFm phases prepared by coprecipitating iodide with other anions were investigated in order to understand this preferential uptake mechanism. X-ray diffraction (XRD) investigations showed a segregation of monoiodide (AFm-I(2)) and Friedel's salt (AFm-Cl(2)) for I-Cl mixtures, whereas interstratifications of AFm-I(2) and hemicarboaluminate (AFm-OH-(CO(3))(0.5)) were observed for the I-CO(3) systems. In contrast, XRD measurements indicated the formation of a solid solution between AFm-I(2) and AFm-SO(4) for the I-SO(4) mixtures. Extended X-ray absorption fine structure spectroscopy showed a modification of the coordination environment of iodine in I-CO(3) and in I-SO(4) samples compared to pure AFm-I(2). This is assumed to be due to the introduction of stacking faults in I-CO(3) samples on one hand and due to the presence of sulfate and associated space-filling water molecules as close neighbors in I-SO(4) samples on the other hand. The formation of a solid solution between AFm-I(2) and AFm-SO(4), with a short-range mixing of iodide and sulfate, implies that AFm-SO(4) bears the potential to retard (129)I. PMID:22376086

  19. [AFM fishing of proteins under impulse electric field].

    PubMed

    Ivanov, Yu D; Pleshakova, T O; Malsagova, K A; Kaysheva, A L; Kopylov, A T; Izotov, A A; Tatur, V Yu; Vesnin, S G; Ivanova, N D; Ziborov, V S; Archakov, A I

    2016-05-01

    A combination of (atomic force microscopy)-based fishing (AFM-fishing) and mass spectrometry allows to capture protein molecules from solutions, concentrate and visualize them on an atomically flat surface of the AFM chip and identify by subsequent mass spectrometric analysis. In order to increase the AFM-fishing efficiency we have applied pulsed voltage with the rise time of the front of about 1 ns to the AFM chip. The AFM-chip was made using a conductive material, highly oriented pyrolytic graphite (HOPG). The increased efficiency of AFM-fishing has been demonstrated using detection of cytochrome b5 protein. Selection of the stimulating pulse with a rise time of 1 ns, corresponding to the GHz frequency range, by the effect of intrinsic emission from water observed in this frequency range during water injection into the cell. PMID:27562998

  20. [AFM fishing of proteins under impulse electric field].

    PubMed

    Ivanov, Yu D; Pleshakova, T O; Malsagova, K A; Kaysheva, A L; Kopylov, A T; Izotov, A A; Tatur, V Yu; Vesnin, S G; Ivanova, N D; Ziborov, V S; Archakov, A I

    2016-05-01

    A combination of (atomic force microscopy)-based fishing (AFM-fishing) and mass spectrometry allows to capture protein molecules from solutions, concentrate and visualize them on an atomically flat surface of the AFM chip and identify by subsequent mass spectrometric analysis. In order to increase the AFM-fishing efficiency we have applied pulsed voltage with the rise time of the front of about 1 ns to the AFM chip. The AFM-chip was made using a conductive material, highly oriented pyrolytic graphite (HOPG). The increased efficiency of AFM-fishing has been demonstrated using detection of cytochrome b5 protein. Selection of the stimulating pulse with a rise time of 1 ns, corresponding to the GHz frequency range, by the effect of intrinsic emission from water observed in this frequency range during water injection into the cell.

  1. Investigation of the heparin-thrombin interaction by dynamic force spectroscopy

    PubMed Central

    Wang, Congzhou; Jin, Yingzi; Desai, Umesh R.; Yadavalli, Vamsi K.

    2015-01-01

    Background The interaction between heparin and thrombin is a vital step in the blood (anti) coagulation process. Unraveling the molecular basis of the interactions is therefore extremely important for understanding the mechanisms of this complex biological process. Methods In this study, we use a combination of an efficient thiolation chemistry of heparin, a self-assembled monolayer-based single molecule platform, and dynamic force spectroscopy to provide new insights into the heparin-thrombin interaction from an energy viewpoint at the molecular scale. Results Well-separated single molecules of heparin covalently attached to mixed self-assembled monolayers are demonstrated, whereby interaction forces with thrombin can be measured via atomic force microscopy-based spectroscopy. Further these interactions are studied at different loading rates and salt concentrations to directly obtain kinetic parameters. Conclusions An increase in the loading rate shows a higher interaction force between the heparin and thrombin, which can be directly linked to the kinetic dissociation rate constant (koff ). The stability of the heparin/thrombin complex decreased with increasing NaCl concentration such that the off-rate was found to be driven primarily by non-ionic forces. General Significance These results contribute to understanding the role of specific and nonspecific forces that drive heparin-thrombin interactions under applied force or flow conditions. PMID:25647100

  2. Direct observation of hydration of TiO 2 on Ti using electrochemical AFM: freely corroding versus potentiostatically held

    NASA Astrophysics Data System (ADS)

    Bearinger, Jane P.; Orme, Christine A.; Gilbert, Jeremy L.

    2001-10-01

    Hydration of titanium/titanium oxide surfaces under freely corroding and potentiostatically held conditions has been characterized using electrochemical atomic force microscopy (EC AFM). In contrast to conventional high vacuum techniques, AFM enables measurement of morphological surface structure in the in situ hydrated state. Electrochemical probes in the imaging environment further enable acquisition of electrical characteristics during AFM imaging. Experiments were performed on etched, electropolished commercially pure titanium. As noted by direct observation and corroborated by power spectral density (Fourier analysis) measurements, oxide domes cover the titanium surface and grow laterally during hydration. Applied potential altered the growth rate. Under open circuit potential conditions, growth proceeded approximately six times faster than under a -1 V applied voltage ( 1098±52 nm2/ min ± versus 184.84±19 nm2/min). Film growth increased electrical resistance and lowered interfacial capacitance based on step polarization impedance spectroscopy tests.

  3. Monitoring TriAcylGlycerols Accumulation by Atomic Force Microscopy Based Infrared Spectroscopy in Streptomyces Species for Biodiesel Applications.

    PubMed

    Deniset-Besseau, Ariane; Prater, Craig B; Virolle, Marie-Joëlle; Dazzi, Alexandre

    2014-02-20

    An atomic force microscope coupled with a tunable infrared laser source (AFM-IR) was used to measure the size and map the distribution of oil inclusions inside of microorganism without staining or other special sample preparation. The microorganism under study is Streptomyces, a soil bacterium that possesses the capability, under some specific nutritional conditions, to store its carbon source into TriAcylGlycerols, a potential direct source of biodiesel. PMID:26270832

  4. Testing synthetic amyloid-β aggregation inhibitor using single molecule atomic force spectroscopy.

    PubMed

    Hane, Francis T; Lee, Brenda Y; Petoyan, Anahit; Rauk, Arvi; Leonenko, Zoya

    2014-04-15

    Alzheimer's disease is a neurodegenerative disease with no known cure and few effective treatment options. The principal neurotoxic agent is an oligomeric form of the amyloid-β peptide and one of the treatment options currently being studied is the inhibition of amyloid aggregation. In this work, we test a novel pseudopeptidic aggregation inhibitor designated as SG1. SG1 has been designed to bind at the amyloid-β self-recognition site and prevent amyloid-β from misfolding into β sheet. We used atomic force spectroscopy, a nanoscale measurement technique, to quantify the binding forces between two single amyloid peptide molecules. For the first time, we demonstrate that single molecule atomic force spectroscopy can be used to assess the effectiveness of amyloid aggregation inhibitors by measuring the experimental yield of binding and can potentially be used as a screening technique for quick testing of efficacy of inhibitor drugs for amyloid aggregation.

  5. High Q optical fiber tips for NC-AFM in liquid.

    PubMed

    LeDue, J M; Lopez-Ayon, M; Burke, S A; Miyahara, Y; Grütter, P

    2009-07-01

    Non-contact atomic force microscopy is rapidly expanding from ultra-high vacuum to include the study of surfaces and biomolecules in liquids by high resolution imaging and force spectroscopy. This is despite the additional frequency shift noise due to the inherently low Q factor of the cantilever oscillating in a liquid. In this paper we present a tip based on an optical fiber which can operate in liquid with Q factors in excess of 100 using a 'diving bell' arrangement which allows only a small portion of the tip to be submerged. We demonstrate stable imaging and force spectroscopy using this set-up. The tips are based on scanning near-field optical microscopy tips and, when used with NC-AFM, provide a method of combining both high resolution mechanical and fluorescence studies of biomolecules and cells. PMID:19509445

  6. Single-molecule force spectroscopy studies of fibrin 'A-a' polymerization interactions via the atomic force microscope

    NASA Astrophysics Data System (ADS)

    Averett, Laurel E.

    Fibrin, the polymerized form of the soluble plasma protein fibrinogen, plays a critical role in hemostasis as the structural scaffold of blood clots. The primary functions of fibrin are to withstand the shear forces of blood flow and provide mechanical stability to the clot, protecting the wound. While studies have investigated the mechanical properties of fibrin constructs, the response to force of critical polymerization interactions such as the 'A--a' knob--hole interaction remains unclear. Herein, the response of the 'A--a' bond to force was examined at the single-molecule level using the atomic force microscope. Force spectroscopy methodology was developed to examine the 'A--a' interaction while reducing the incidence of both nonspecific and multiple molecule interactions. The rupture of this interaction resulted in a previously unreported characteristic force profile comprised of up to four events. We hypothesized that the first event represented reorientation of the fibrinogen molecule, the second and third represented unfolding of structures in the D region of fibrinogen, and the last event was the rupture of the 'A--a' bond weakened by prior structural unfolding. The configuration, molecular extension, and kinetic parameters of each event in the characteristic pattern were examined to compare the unfolding of fibrin to other proteins unfolded by force. Fitting the pattern with polymer models showed that the D region of fibrinogen could lengthen by ˜50% of the length of a fibrin monomer before rupture of the 'A--a' bond. Analysis showed that the second and third events had kinetic parameters similar to other protein structures unfolded by force. Studies of the dependence of the characteristic pattern on calcium, concentration of sodium chloride, pH, and temperature demonstrated that the incidence of the last event was affected by solution conditions. However, only low pH and high temperatures reduced the probability that an interaction was characteristic

  7. Dynamic force spectroscopy of the Helicobacter pylori BabA-Lewis b binding.

    PubMed

    Björnham, Oscar; Bugaytsova, Jeanna; Borén, Thomas; Schedin, Staffan

    2009-07-01

    The binding strength of the Helicobacter pylori adhesin-receptor complex BabA-ABO/Lewis b has been analyzed by means of dynamic force spectroscopy. High-resolution measurements of rupture forces were performed in situ on single bacterial cells, expressing the high-affinity binding BabA adhesin, by the use of force measuring optical tweezers. The resulting force spectra revealed the mechanical properties of a single BabA-Leb bond. It was found that the bond is dominated by one single energy barrier and that it is a slip-bond. The bond length and thermal off-rate were assessed to be 0.86+/-0.07 nm and 0.015+/-0.006 s(-1), respectively.

  8. Assembly of live micro-organisms on microstructured PDMS stamps by convective/capillary deposition for AFM bio-experiments.

    PubMed

    Dague, E; Jauvert, E; Laplatine, L; Viallet, B; Thibault, C; Ressier, L

    2011-09-30

    Immobilization of live micro-organisms on solid substrates is an important prerequisite for atomic force microscopy (AFM) bio-experiments. The method employed must immobilize the cells firmly enough to enable them to withstand the lateral friction forces exerted by the tip during scanning but without denaturing the cell interface. In this work, a generic method for the assembly of living cells on specific areas of substrates is proposed. It consists in assembling the living cells within the patterns of microstructured, functionalized poly-dimethylsiloxane (PDMS) stamps using convective/capillary deposition. This versatile approach is validated by applying it to two systems of foremost importance in biotechnology and medicine: Saccharomyces cerevisiae yeasts and Aspergillus fumigatus fungal spores. We show that this method allows multiplexing AFM nanomechanical measurements by force spectroscopy on S. cerevisiae yeasts and high-resolution AFM imaging of germinated Aspergillus conidia in buffer medium. These two examples clearly demonstrate the immense potential of micro-organism assembly on functionalized, microstructured PDMS stamps by convective/capillary deposition for performing rigorous AFM bio-experiments on living cells.

  9. Assembly of live micro-organisms on microstructured PDMS stamps by convective/capillary deposition for AFM bio-experiments

    NASA Astrophysics Data System (ADS)

    Dague, E.; Jauvert, E.; Laplatine, L.; Viallet, B.; Thibault, C.; Ressier, L.

    2011-09-01

    Immobilization of live micro-organisms on solid substrates is an important prerequisite for atomic force microscopy (AFM) bio-experiments. The method employed must immobilize the cells firmly enough to enable them to withstand the lateral friction forces exerted by the tip during scanning but without denaturing the cell interface. In this work, a generic method for the assembly of living cells on specific areas of substrates is proposed. It consists in assembling the living cells within the patterns of microstructured, functionalized poly-dimethylsiloxane (PDMS) stamps using convective/capillary deposition. This versatile approach is validated by applying it to two systems of foremost importance in biotechnology and medicine: Saccharomyces cerevisiae yeasts and Aspergillus fumigatus fungal spores. We show that this method allows multiplexing AFM nanomechanical measurements by force spectroscopy on S. cerevisiae yeasts and high-resolution AFM imaging of germinated Aspergillus conidia in buffer medium. These two examples clearly demonstrate the immense potential of micro-organism assembly on functionalized, microstructured PDMS stamps by convective/capillary deposition for performing rigorous AFM bio-experiments on living cells.

  10. Surface morphology and chemistry of Prunus laurocerasus L. leaves: a study using X-ray photoelectron spectroscopy, time-of-flight secondary-ion mass spectrometry, atomic-force microscopy and scanning-electron microscopy.

    PubMed

    Perkins, Mark C; Roberts, Clive J; Briggs, David; Davies, Martyn C; Friedmann, Adrian; Hart, Clifford A; Bell, Gordon A

    2005-04-01

    The surface properties of the plant cuticle play a crucial role in plant-pathogen interactions and the retention and penetration of agriculturally important chemicals. This paper describes the use of X-ray photoelectron spectroscopy (XPS), time-of-flight secondary-ion mass spectrometry (ToF-SIMS), tapping-mode atomic force microscopy (TM-AFM) and scanning electron microscopy (SEM) to determine surface-specific chemical and material properties of the adaxial surface of Prunus laurocerasus L. leaves. XPS data, derived from the uppermost few nanometres (< 10 nm) of the leaf surface, were consistent with the wax components and functionality known to be present within the waxes. ToF-SIMS provided molecular speciation from the outermost monolayer of the leaf surface, indicating the importance of a family of acetates with chain lengths ranging from C20 to C34. The presence of alkanes with C29 and C31 chain lengths was also confirmed. SEM and TM-AFM topography images revealed a textured granular surface, while simultaneously recorded AFM phase images revealed heterogeneous material properties at the nanoscale. The relevance of these data to plant cuticle development, allelochemistry and agrochemical delivery is discussed.

  11. Detecting cell-adhesive sites in extracellular matrix using force spectroscopy mapping

    PubMed Central

    Chirasatitsin, Somyot; Engler, Adam J

    2010-01-01

    The cell microenvironment is composed of extracellular matrix (ECM), which contains specific binding sites that allow the cell to adhere to its surroundings. Cells employ focal adhesion proteins, which must be able to resist a variety of forces to bind to ECM. Current techniques for detecting the spatial arrangement of these adhesions, however, have limited resolution and those that detect adhesive forces lack sufficient spatial characterization or resolution. Using a unique application of force spectroscopy, we demonstrate here the ability to determine local changes in the adhesive property of a fibronectin substrate down to the resolution of the fibronectin antibody-functionalized tip diameter, ~20 nm. To verify the detection capabilities of force spectroscopy mapping (FSM), changes in loading rate and temperature were used to alter the bond dynamics and change the adhesion force. Microcontact printing was also used to pattern fluorescein isothiocyanate-conjugated fibronectin in order to mimic the discontinuous adhesion domains of native ECM. Fluorescent detection was used to identify the pattern while FSM was used to map cell adhesion sites in registry with the initial fluorescent image. The results show that FSM can be used to detect the adhesion domains at high resolution and may subsequently be applied to native ECM with randomly distributed cell adhesion sites. PMID:21152375

  12. Molecular force modulation spectroscopy revealing the dynamic response of single bacteriorhodopsins.

    PubMed

    Janovjak, Harald; Müller, Daniel J; Humphris, Andrew D L

    2005-02-01

    Recent advances in atomic force microscopy allowed globular and membrane proteins to be mechanically unfolded on a single-molecule level. Presented is an extension to the existing force spectroscopy experiments. While unfolding single bacteriorhodopsins from native purple membranes, small oscillation amplitudes (6-9 nm) were supplied to the vertical displacement of the cantilever at a frequency of 3 kHz. The phase and amplitude response of the cantilever-protein system was converted to reveal the elastic (conservative) and viscous (dissipative) contributions to the unfolding process. The elastic response (stiffness) of the extended parts of the protein were in the range of a few tens pN/nm and could be well described by the derivative of the wormlike chain model. Discrete events in the viscous response coincided with the unfolding of single secondary structure elements and were in the range of 1 microNs/m. In addition, these force modulation spectroscopy experiments revealed novel mechanical unfolding intermediates of bacteriorhodopsin. We found that kinks result in a loss of unfolding cooperativity in transmembrane helices. Reconstructing force-distance spectra by the integration of amplitude-distance spectra verified their position, offering a novel approach to detect intermediates during the forced unfolding of single proteins. PMID:15574708

  13. Nanomechanical characterization of phospholipid bilayer islands on flat and porous substrates: a force spectroscopy study.

    PubMed

    Nussio, Matthew R; Oncins, Gerard; Ridelis, Ingrid; Szili, Endre; Shapter, Joseph G; Sanz, Fausto; Voelcker, Nicolas H

    2009-07-30

    In this study, we compare for the first time the nanomechanical properties of lipid bilayer islands on flat and porous surfaces. 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) bilayers were deposited on flat (silicon and mica) and porous silicon (pSi) substrate surfaces and examined using atomic force spectroscopy and force volume imaging. Force spectroscopy measurements revealed the effects of the underlying substrate and of the lipid phase on the nanomechanical properties of bilayers islands. For mica and silicon, significant differences in breakthrough force between the center and the edges of bilayer islands were observed for both phospolipids. These differences were more pronounced for DMPC than for DPPC, presumably due to melting effects at the edges of DMPC bilayers. In contrast, bilayer islands deposited on pSi yielded similar breakthrough forces in the central region and along the perimeter of the islands, and those values in turn were similar to those measured along the perimeter of bilayer islands deposited on the flat substrates. The study also demonstrates that pSi is suitable solid support for the formation of pore-spanning phospholipid bilayers with potential applications in transmembrane protein studies, drug delivery, and biosensing.

  14. Crystallographic order and decomposition of [MnIII6CrIII]3+ single-molecule magnets deposited in submonolayers and monolayers on HOPG studied by means of molecular resolved atomic force microscopy (AFM) and Kelvin probe force microscopy in UHV.

    PubMed

    Gryzia, Aaron; Volkmann, Timm; Brechling, Armin; Hoeke, Veronika; Schneider, Lilli; Kuepper, Karsten; Glaser, Thorsten; Heinzmann, Ulrich

    2014-01-01

    Monolayers and submonolayers of [MnIII6CrIII]3+ single-molecule magnets (SMMs) adsorbed on highly oriented pyrolytic graphite (HOPG) using the droplet technique characterized by non-contact atomic force microscopy (nc-AFM) as well as by Kelvin probe force microscopy (KPFM) show island-like structures with heights resembling the height of the molecule. Furthermore, islands were found which revealed ordered 1D as well as 2D structures with periods close to the width of the SMMs. Along this, islands which show half the heights of intact SMMs were observed which are evidences for a decomposing process of the molecules during the preparation. Finally, models for the structure of the ordered SMM adsorbates are proposed to explain the observations.

  15. Characterization of actomyosin bond properties in intact skeletal muscle by force spectroscopy

    PubMed Central

    Colombini, Barbara; Bagni, M. Angela; Romano, Giovanni; Cecchi, Giovanni

    2007-01-01

    Force generation and motion in skeletal muscle result from interaction between actin and myosin myofilaments through the cyclical formation and rupture of the actomyosin bonds, the cross-bridges, in the overlap region of the sarcomeres. Actomyosin bond properties were investigated here in single intact muscle fibers by using dynamic force spectroscopy. The force needed to forcibly detach the cross-bridge ensemble in the half-sarcomere (hs) was measured in a range of stretching velocity between 3.4 × 103 nm·hs−1·s−1 or 3.3 fiber length per second (l0s−1) and 6.1 × 104 nm·hs−1·s−1 or 50 l0·s−1 during tetanic force development. The rupture force of the actomyosin bond increased linearly with the logarithm of the loading rate, in agreement with previous experiments on noncovalent single bond and with Bell theory [Bell GI (1978) Science 200:618–627]. The analysis permitted calculation of the actomyosin interaction length, xβ and the dissociation rate constant for zero external load, k0. Mean xβ was 1.25 nm, a value similar to that reported for single actomyosin bond under rigor condition. Mean k0 was 20 s−1, a value about twice as great as that reported in the literature for isometric force relaxation in the same type of muscle fibers. These experiments show, for the first time, that force spectroscopy can be used to reveal the properties of the individual cross-bridge in intact skeletal muscle fibers. PMID:17517641

  16. Contact nanomechanical measurements with the AFM

    NASA Astrophysics Data System (ADS)

    Geisse, Nicholas

    2013-03-01

    The atomic force microscope (AFM) has found broad use in the biological sciences largely due to its ability to make measurements on unfixed and unstained samples under liquid. In addition to imaging at multiple spatial scales ranging from micro- to nanometer, AFMs are commonly used as nanomechanical probes. This is pertinent for cell biology, as it has been demonstrated that the geometrical and mechanical properties of the extracellular microenvironment are important in such processes as cancer, cardiovascular disease, muscular dystrophy, and even the control of cell life and death. Indeed, the ability to control and quantify these external geometrical and mechanical parameters arises as a key issue in the field. Because AFM can quantitatively measure the mechanical properties of various biological samples, novel insights to cell function and to cell-substrate interactions are now possible. As the application of AFM to these types of problems is widened, it is important to understand the performance envelope of the technique and its associated data analyses. This talk will discuss the important issues that must be considered when mechanical models are applied to real-world data. Examples of the effect of different model assumptions on our understanding of the measured material properties will be shown. Furthermore, specific examples of the importance of mechanical stimuli and the micromechanical environment to the structure and function of biological materials will be presented.

  17. AFM CHARACTERIZATION OF RAMAN LASER INDUCED DAMAGE ON CDZNTECRYSTAL SURFACES

    SciTech Connect

    Teague, L.; Duff, M.

    2008-10-07

    High quality CdZnTe (or CZT) crystals have the potential for use in room temperature gamma-ray and X-ray spectrometers. Over the last decade, the methods for growing high quality CZT have improved the quality of the produced crystals however there are material features that can influence the performance of these materials as radiation detectors. The presence of structural heterogeneities within the crystals, such as twinning, pipes, grain boundaries (polycrystallinity), and secondary phases (SPs) can have an impact on the detector performance. There is considerable need for reliable and reproducible characterization methods for the measurement of crystal quality. With improvements in material characterization and synthesis, these crystals may become suitable for widespread use in gamma radiation detection. Characterization techniques currently utilized to test for quality and/or to predict performance of the crystal as a gamma-ray detector include infrared (IR) transmission imaging, synchrotron X-ray topography, photoluminescence spectroscopy, transmission electron microscopy (TEM), atomic force microscopy (AFM) and Raman spectroscopy. In some cases, damage caused by characterization methods can have deleterious effects on the crystal performance. The availability of non-destructive analysis techniques is essential to validate a crystal's quality and its ability to be used for either qualitative or quantitative gamma-ray or X-ray detection. The work presented herein discusses the damage that occurs during characterization of the CZT surface by a laser during Raman spectroscopy, even at minimal laser powers. Previous Raman studies have shown that the localized annealing from tightly focused, low powered lasers results in areas of higher Te concentration on the CZT surface. This type of laser damage on the surface resulted in decreased detector performance which was most likely due to increased leakage current caused by areas of higher Te concentration. In this study

  18. Quantitatively Resolving Ligand–Receptor Bonds on Cell Surfaces Using Force-Induced Remnant Magnetization Spectroscopy

    PubMed Central

    2016-01-01

    Molecule-specific noncovalent bonding on cell surfaces is the foundation for cellular recognition and functioning. A major challenge in probing these bonds is to resolve the specific bonds quantitatively and efficiently from the nonspecific interactions in a complex environment. Using force-induced remnant magnetization spectroscopy (FIRMS), we were able to resolve quantitatively three different interactions for magnetic beads bearing anti-CD4 antibodies with CD4+ T cell surfaces based upon their binding forces. The binding force of the CD4 antibody–antigen bonds was determined to be 75 ± 3 pN. For comparison, the same bonds were also studied on a functionalized substrate surface, and the binding force was determined to be 90 ± 6 pN. The 15 pN difference revealed by high-resolution FIRMS illustrates the significant impact of the bonding environment. Because the force difference was unaffected by the cell number or the receptor density on the substrate, we attributed it to the possible conformational or local environmental differences of the CD4 antigens between the cell surface and substrate surface. Our results show that the high force resolution and detection efficiency afforded by FIRMS are valuable for studying protein–protein interactions on cell surfaces. PMID:27163031

  19. Hydrophobic interaction governs unspecific adhesion of staphylococci: a single cell force spectroscopy study.

    PubMed

    Thewes, Nicolas; Loskill, Peter; Jung, Philipp; Peisker, Henrik; Bischoff, Markus; Herrmann, Mathias; Jacobs, Karin

    2014-01-01

    Unspecific adhesion of bacteria is usually the first step in the formation of biofilms on abiotic surfaces, yet it is unclear up to now which forces are governing this process. Alongside long-ranged van der Waals and electrostatic forces, short-ranged hydrophobic interaction plays an important role. To characterize the forces involved during approach and retraction of an individual bacterium to and from a surface, single cell force spectroscopy is applied: A single cell of the apathogenic species Staphylococcus carnosus isolate TM300 is used as bacterial probe. With the exact same bacterium, hydrophobic and hydrophilic surfaces can be probed and compared. We find that as far as 50 nm from the surface, attractive forces can already be recorded, an indication of the involvement of long-ranged forces. Yet, comparing the surfaces of different surface energy, our results corroborate the model that large, bacterial cell wall proteins are responsible for adhesion, and that their interplay with the short-ranged hydrophobic interaction of the involved surfaces is mainly responsible for adhesion. The ostensibly long range of the attraction is a result of the large size of the cell wall proteins, searching for contact via hydrophobic interaction. The model also explains the strong (weak) adhesion of S. carnosus to hydrophobic (hydrophilic) surfaces. PMID:25247133

  20. Investigation of free fatty acid associated recombinant membrane receptor protein expression in HEK293 cells using Raman spectroscopy, calcium imaging, and atomic force microscopy.

    PubMed

    Lin, Juqiang; Xu, Han; Wu, Yangzhe; Tang, Mingjie; McEwen, Gerald D; Liu, Pin; Hansen, Dane R; Gilbertson, Timothy A; Zhou, Anhong

    2013-02-01

    G-protein-coupled receptor 120 (GPR120) is a previously orphaned G-protein-coupled receptor that apparently functions as a sensor for dietary fat in the gustatory and digestive systems. In this study, a cDNA sequence encoding a doxycycline (Dox)-inducible mature peptide of GPR120 was inserted into an expression vector and transfected in HEK293 cells. We measured Raman spectra of single HEK293 cells as well as GPR120-expressing HEK293-GPR120 cells at a 48 h period following the additions of Dox at several concentrations. We found that the spectral intensity of HEK293-GPR120 cells is dependent upon the dose of Dox, which correlates with the accumulation of GPR120 protein in the cells. However, the amount of the fatty acid activated changes in intracellular calcium (Ca(2+)) as measured by ratiometric calcium imaging was not correlated with Dox concentration. Principal components analysis (PCA) of Raman spectra reveals that the spectra from different treatments of HEK293-GPR120 cells form distinct, completely separated clusters with the receiver operating characteristic (ROC) area of 1, while those spectra for the HEK293 cells form small overlap clusters with the ROC area of 0.836. It was also found that expression of GPR120 altered the physiochemical and biomechanical properties of the parental cell membrane surface, which was quantitated by atomic force microscopy (AFM). These findings demonstrate that the combination of Raman spectroscopy, calcium imaging, and AFM may provide new tools in noninvasive and quantitative monitoring of membrane receptor expression induced alterations in the biophysical and signaling properties of single living cells.

  1. Directly measuring single molecule heterogeneity in proteins and RNA using force spectroscopy

    NASA Astrophysics Data System (ADS)

    Hinczewski, Michael; Hyeon, Changbong; Thirumalai, Devarajan

    One of the most intriguing results of single molecule experiments on proteins and nucleic acids is the discovery of functional heterogeneity: the observation that complex cellular machines exhibit multiple, biologically active conformations. The structural differences between these conformations may be subtle, but each distinct state can be remarkably long-lived, with stochastic interconversions occurring only at macroscopic timescales, fractions of a second or longer. Though we now have proof of functional heterogeneity in a handful of systems--enzymes, motors, adhesion complexes--identifying and measuring it remains a formidable challenge. We show that evidence of this phenomenon is more widespread than previously known, encoded in data collected from some of the most well-established single molecule techniques: AFM or optical tweezer pulling experiments. We present a theoretical procedure for analyzing distributions of rupture/unfolding forces recorded at different pulling speeds. This results in a single parameter, quantifying the degree of heterogeneity, and also leads to bounds on the equilibration and conformational interconversion timescales. Our work suggests experimental approaches for estimating the timescales of these fluctuations with unprecedented accuracy.

  2. Rational fabrication of a gold-coated AFM TERS tip by pulsed electrodeposition.

    PubMed

    Yang, Li-Kun; Huang, Teng-Xiang; Zeng, Zhi-Cong; Li, Mao-Hua; Wang, Xiang; Yang, Fang-Zu; Ren, Bin

    2015-11-21

    Reproducible fabrication of sharp gold- or silver-coated tips has become the bottleneck issue in tip-enhanced Raman spectroscopy, especially for atomic force microscopy (AFM)-based TERS. Herein, we developed a novel method based on pulsed electrodeposition to coat a thin gold layer over atomic force microscopy (AFM) tips to produce plasmonic TERS tips with high reproducibility. We systematically investigated the influence of the deposition potential and step time on the surface roughness and sharpness. This method allows the rational control of the radii of gold-coated TERS tips from a few to hundreds of nanometers, which allows us to systematically study the dependence of the TERS enhancement on the radius of the gold-coated AFM tip. The maximum TERS enhancement was achieved for the tip radius in the range of 60-75 nm in the gap mode. The coated gold layer has a strong adhesion with the silicon tip surface, which is highly stable in water, showing the great potential for application in the aqueous environment. PMID:26482226

  3. Harmonic force spectroscopy reveals a force-velocity curve from a single human beta cardiac myosin motor

    NASA Astrophysics Data System (ADS)

    Sung, Jongmin; Nag, Suman; Vestergaard, Christian; Mortensen, Kim; Flyvbjerg, Henrik; Spudich, James

    2014-03-01

    A muscle contracts rapidly under low load, but slowly under high load. Its molecular mechanisms remain to be elucidated, however. During contraction, myosins in thick filaments interact with actin in thin filaments in the sarcomere, cycling between a strongly bound (force producing) state and a weakly bound (relaxed) state. Huxley et al. have previously proposed that the transition from the strong to the weak interaction can be modulated by a load. We use a new method we call ``harmonic force spectroscopy'' to extract a load-velocity curve from a single human beta cardiac myosin II motor. With a dual-beam optical trap, we hold an actin dumbbell over a myosin molecule anchored to the microscope stage that oscillates sinusoidally. Upon binding, the motor experiences an oscillatory load with a mean that is directed forward or backward, depending on binding location We find that the bound time at saturating [ATP] is exponentially correlated with the mean load, which is explained by Arrhenius transition theory. With a stroke size measurement, we obtained a load-velocity curve from a single myosin. We compare the curves for wild-type motors with mutants that cause hypertrophic cardiomyopathies, to understand the effects on the contractile cycle

  4. A single-molecule force spectroscopy study of the interactions between lectins and carbohydrates on cancer and normal cells

    NASA Astrophysics Data System (ADS)

    Zhao, Weidong; Cai, Mingjun; Xu, Haijiao; Jiang, Junguang; Wang, Hongda

    2013-03-01

    The interaction forces between carbohydrates and lectins were investigated by single-molecule force spectroscopy on both cancer and normal cells. The binding kinetics was also studied, which shows that the carbohydrate-lectin complex on cancer cells is less stable than that on normal cells.The interaction forces between carbohydrates and lectins were investigated by single-molecule force spectroscopy on both cancer and normal cells. The binding kinetics was also studied, which shows that the carbohydrate-lectin complex on cancer cells is less stable than that on normal cells. Electronic supplementary information (ESI) available: Experimental details. See DOI: 10.1039/c3nr00553d

  5. Raman and AFM study of gamma irradiated plastic bottle sheets

    NASA Astrophysics Data System (ADS)

    Ali, Yasir; Kumar, Vijay; Sonkawade, R. G.; Dhaliwal, A. S.

    2013-02-01

    In this investigation, the effects of gamma irradiation on the structural properties of plastic bottle sheet are studied. The Plastic sheets were exposed with 1.25MeV 60Co gamma rays source at various dose levels within the range from 0-670 kGy. The induced modifications were followed by micro-Raman and atomic force microscopy (AFM). The Raman spectrum shows the decrease in Raman intensity and formation of unsaturated bonds with an increase in the gamma dose. AFM image displays rough surface morphology after irradiation. The detailed Raman analysis of plastic bottle sheets is presented here, and the results are correlated with the AFM observations.

  6. Raman and AFM study of gamma irradiated plastic bottle sheets

    SciTech Connect

    Ali, Yasir; Kumar, Vijay; Dhaliwal, A. S.; Sonkawade, R. G.

    2013-02-05

    In this investigation, the effects of gamma irradiation on the structural properties of plastic bottle sheet are studied. The Plastic sheets were exposed with 1.25MeV {sup 60}Co gamma rays source at various dose levels within the range from 0-670 kGy. The induced modifications were followed by micro-Raman and atomic force microscopy (AFM). The Raman spectrum shows the decrease in Raman intensity and formation of unsaturated bonds with an increase in the gamma dose. AFM image displays rough surface morphology after irradiation. The detailed Raman analysis of plastic bottle sheets is presented here, and the results are correlated with the AFM observations.

  7. Single-cell force spectroscopy of the medically important Staphylococcus epidermidis-Candida albicans interaction

    NASA Astrophysics Data System (ADS)

    Beaussart, Audrey; Herman, Philippe; El-Kirat-Chatel, Sofiane; Lipke, Peter N.; Kucharíková, Soňa; van Dijck, Patrick; Dufrêne, Yves F.

    2013-10-01

    Despite the clinical importance of bacterial-fungal interactions, their molecular details are poorly understood. A hallmark of such medically important interspecies associations is the interaction between the two nosocomial pathogens Staphylococcus aureus and Candida albicans, which can lead to mixed biofilm-associated infections with enhanced antibiotic resistance. Here, we use single-cell force spectroscopy (SCFS) to quantify the forces engaged in bacterial-fungal co-adhesion, focusing on the poorly investigated S. epidermidis-C. albicans interaction. Force curves recorded between single bacterial and fungal germ tubes showed large adhesion forces (~5 nN) with extended rupture lengths (up to 500 nm). By contrast, bacteria poorly adhered to yeast cells, emphasizing the important role of the yeast-to-hyphae transition in mediating adhesion to bacterial cells. Analysis of mutant strains altered in cell wall composition allowed us to distinguish the main fungal components involved in adhesion, i.e. Als proteins and O-mannosylations. We suggest that the measured co-adhesion forces are involved in the formation of mixed biofilms, thus possibly as well in promoting polymicrobial infections. In the future, we anticipate that this SCFS platform will be used in nanomedicine to decipher the molecular mechanisms of a wide variety of pathogen-pathogen interactions and may help in designing novel anti-adhesion agents.

  8. Single-cell force spectroscopy of the medically-important Staphyloccocus epidermidis-Candida albicans interaction

    PubMed Central

    Beaussart, Audrey; Herman, Philippe; El-Kirat-Chatel, Sofiane; Lipke, Peter N.; Kucharíková, Soňa; Van Dijck, Patrick; Dufrêne, Yves F.

    2013-01-01

    Despite the clinical importance of bacterial-fungal interactions, their molecular details are poorly understood. A hallmark of such medically-important interspecies associations is the interaction between the two nosocomial pathogens Staphylococcus aureus and Candida albicans, which can lead to mixed biofilm-associated infections with enhanced antibiotic resistance. Here, we use single-cell force spectroscopy to quantify the forces engaged in bacterial-fungal co-adhesion, focusing on the poorly investigated S. epidermidis-C. albicans interaction. Force curves recorded between single bacterial and fungal germ tubes showed large adhesion forces (~5 nN) with extended rupture lengths (up to 500 nm). By contrast, bacteria poorly adhered to yeast cells, emphasizing the important role of the yeast-to-hyphae transition in mediating adhesion to bacterial cells. Analysis of mutant strains altered in cell wall composition allowed us to distinguish the main fungal components involved in adhesion, i.e. Als proteins and O-mannosylations. We suggest that the measured co-adhesion forces are involved in the formation of mixed biofilms, thus possibly as well in promoting polymicrobial infections. In the future, we anticipate that this SCFS platform will be used in nanomedicine to decipher the molecular mechanisms of a wide variety of pathogen-pathogen interactions and may help designing novel anti-adhesion agents. PMID:24057018

  9. Force spectroscopy of DNA: there is still a lot to learn

    NASA Astrophysics Data System (ADS)

    Paik, D. H.; Perkins, Thomas T.

    2012-10-01

    Single-molecule studies of the mechanical properties of individual double-stranded DNA have excited interest across many scientific disciplines because of DNA's fundamental role in biology and DNA's remarkable overstretching transition at higher forces. Here, we discuss a recent result on the overstretching transition of DNA and on the dynamics of dye molecules intercalating into DNA under tension. Overstretching DNA is mechanical transition whereby DNA's extension increases by 70% at 65 pN. Notwithstanding more than a decade of experimental and theoretical studies, there remains significant debate on the nature of overstretched DNA. We developed a topologically closed but torsionally unconstrained DNA assay that contains no nicks or free ends. DNA in this assay exhibited the canonical overstretching transition at 65 pN but without hysteresis upon retraction. Controlled introduction of a nick led to hysteresis in the force extension curve. Moreover, the degree of hysteresis increased with the number of nicks. In the second study, we isolated the effects of binding and intercalation of a DNA staining dye, by combining single molecule force spectroscopy with simple buffer exchange. We showed that force-enhanced intercalation can occur from a reservoir of bound dye that was not bis-intercalated, yet remained out of equilibrium with free dye for long periods (<5 min for YOPRO and <2 hr for YOYO). Our work highlights that binding/unbinding and intercalation/de-intercalation are distinct processes that can occur on very different time scales. Taken together, these works highlight ongoing discoveries based on a twenty year old technique, force spectroscopy of single DNA molecules.

  10. Dielectrophoretic Tweezers as a Platform for Single Molecular Force Spectroscopy in a Highly Parallel Format

    NASA Astrophysics Data System (ADS)

    Cheng, Peng; Barrett, Michael; Oliver, Piercen; Vezenov, Dmitri

    2012-02-01

    Miniaturization has driven down the cost of tools used in bioanalysis and diagnostics, with single molecules becoming the ultimate detection limit. I will describe how one can exploit mechanical properties of individual biomolecules to determine changes in their state or structure. Our aim is to build a force-spectroscopy-on-a-chip device that can detect and manipulate many (millions) single molecules in parallel. A critical element of this approach is the design of materials properties of molecular handles or probes. By tuning interactions of these probes with electric fields which generate by a simple electrode geometry, we are able to apply piconewton forces to individual DNA molecules and record their response with a single base sensitivity. I will present how we determined the approximate crossover frequency between negative and positive DEP using plain electrodes instead of conventional micro-structures. The technique is attractive not only for conducting single molecule force spectroscopy but also for label-free single cell detection. I will discuss potential applications of this approach to high throughput analyses such as genome sequencing and HIV detection.

  11. Nanomedicine: AFM tackles osteoarthritis

    NASA Astrophysics Data System (ADS)

    Aigner, Thomas; Schmitz, Nicole; Haag, Jochen

    2009-03-01

    Current diagnostic tools detect cartilage degeneration only at advanced stages, but the atomic force microscope can now detect structural changes earlier, paving the way for treatment of joint diseases.

  12. AFM nanoindentations of diatom biosilica surfaces.

    PubMed

    Losic, Dusan; Short, Ken; Mitchell, James G; Lal, Ratnesh; Voelcker, Nicolas H

    2007-04-24

    Diatoms have intricately and uniquely nanopatterned silica exoskeletons (frustules) and are a common target of biomimetic investigations. A better understanding of the diatom frustule structure and function at the nanoscale could provide new insights for the biomimetic fabrication of nanostructured ceramic materials and lightweight, yet strong, scaffold architectures. Here, we have mapped the nanoscale mechanical properties of Coscinodiscus sp. diatoms using atomic force microscopy (AFM)-based nanoindentation. Mechanical properties were correlated with the frustule structures obtained from high-resolution AFM and scanning electron microscopy (SEM). Significant differences in the micromechanical properties for the different frustule layers were observed. A comparative study of other related inorganic material including porous silicon films and free-standing membranes as well as porous alumina was also undertaken.

  13. Detection of Pathogens Using AFM and SPR

    NASA Astrophysics Data System (ADS)

    Vaseashta, Ashok

    2005-03-01

    A priori detection of pathogens in food and water has become a subject of paramount importance. Several recent incidents have resulted in the government passing stringent regulations for tolerable amounts of contamination of food products. Identification and/or monitoring of bacterial contamination in food are critical. The conventional methods of pathogen detection require time-consuming steps to arrive disembark at meaningful measurement in a timely manner as the detection time exceeds the time in which perishable food recycles through the food chain distribution. The aim of this presentation is to outline surface plasmon resonance (SPR) and atomic force microscopy (AFM) as two methods for fast detect6ion of pathogens. Theoretical basis of SPR and experimental results of SPR and AFM on E. coli O157:H7 and prion are presented.

  14. Elastic Properties of Nucleic Acids by Single-Molecule Force Spectroscopy.

    PubMed

    Camunas-Soler, Joan; Ribezzi-Crivellari, Marco; Ritort, Felix

    2016-07-01

    We review the current knowledge on the use of single-molecule force spectroscopy techniques to extrapolate the elastic properties of nucleic acids. We emphasize the lesser-known elastic properties of single-stranded DNA. We discuss the importance of accurately determining the elastic response in pulling experiments, and we review the simplest models used to rationalize the experimental data as well as the experimental approaches used to pull single-stranded DNA. Applications used to investigate DNA conformational transitions and secondary structure formation are also highlighted. Finally, we provide an overview of the effects of salt and temperature and briefly discuss the effects of contour length and sequence dependence. PMID:27145878

  15. Elastic Properties of Nucleic Acids by Single-Molecule Force Spectroscopy.

    PubMed

    Camunas-Soler, Joan; Ribezzi-Crivellari, Marco; Ritort, Felix

    2016-07-01

    We review the current knowledge on the use of single-molecule force spectroscopy techniques to extrapolate the elastic properties of nucleic acids. We emphasize the lesser-known elastic properties of single-stranded DNA. We discuss the importance of accurately determining the elastic response in pulling experiments, and we review the simplest models used to rationalize the experimental data as well as the experimental approaches used to pull single-stranded DNA. Applications used to investigate DNA conformational transitions and secondary structure formation are also highlighted. Finally, we provide an overview of the effects of salt and temperature and briefly discuss the effects of contour length and sequence dependence.

  16. Investigations of the intermolecular forces between RDX and polyethylene by force-distance spectroscopy and molecular dynamics simulations.

    PubMed

    Taylor, D E; Strawhecker, K E; Shanholtz, E R; Sorescu, D C; Sausa, R C

    2014-07-10

    The development of novel nanoenergetic materials with enhanced bulk properties requires an understanding of the intermolecular interactions occurring between molecular components. We investigate the surface interactions between 1,3,5-trinitro-1,3,5-triazacyclohexane (RDX) and polyethylene (PE) crystals on the basis of combined use of molecular dynamics (MD) simulations and force-distance spectroscopy, in conjunction with Lifshitz macroscopic theory of van der Waals forces between continuous materials. The binding energy in the RDX-PE system depends both on the degree of PE crystallinity and on the RDX crystal face. Our MD simulations yield binding energies of approximately 132 and 120 mJ/m(2) for 100% amorphous and 100% crystalline PE on RDX (210), respectively. The average value is about 36% greater than our experimental value of 81 ± 15 mJ/m(2) for PE (∼48% amorphous) on RDX (210). By comparison, Liftshitz theory predicts a value of about 79 mJ/m(2) for PE interacting with RDX. Our MD simulations also predict larger binding energies for both amorphous and crystalline PE on RDX (210) compared to the RDX (001) surface. Analysis of the interaction potential indicates that about 60% of the binding energy in the PE-RDX system is due to attractive interactions between HPE-ORDX and CPE-NRDX pairs of atoms. Further, amorphous PE shows a much longer interaction distance than crystalline PE with the (210) and (001) RDX surfaces due to the possibility of larger polymer elongations in the case of amorphous PE as strain is applied. Also, we report estimates of the binding energies of energetic materials RDX and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) with PE, propylene, polystyrene, and several fluorine-containing polymers using Lifshitz theory and compare these with reported MD calculations.

  17. Harmonic force spectroscopy measures load-dependent kinetics of individual human β-cardiac myosin molecules

    NASA Astrophysics Data System (ADS)

    Sung, Jongmin; Nag, Suman; Mortensen, Kim I.; Vestergaard, Christian L.; Sutton, Shirley; Ruppel, Kathleen; Flyvbjerg, Henrik; Spudich, James A.

    2015-08-01

    Molecular motors are responsible for numerous cellular processes from cargo transport to heart contraction. Their interactions with other cellular components are often transient and exhibit kinetics that depend on load. Here, we measure such interactions using `harmonic force spectroscopy'. In this method, harmonic oscillation of the sample stage of a laser trap immediately, automatically and randomly applies sinusoidally varying loads to a single motor molecule interacting with a single track along which it moves. The experimental protocol and the data analysis are simple, fast and efficient. The protocol accumulates statistics fast enough to deliver single-molecule results from single-molecule experiments. We demonstrate the method's performance by measuring the force-dependent kinetics of individual human β-cardiac myosin molecules interacting with an actin filament at physiological ATP concentration. We show that a molecule's ADP release rate depends exponentially on the applied load, in qualitative agreement with cardiac muscle, which contracts with a velocity inversely proportional to external load.

  18. AFM single-cell force spectroscopy links altered nuclear and cytoskeletal mechanics to defective cell adhesion in cardiac myocytes with a nuclear lamin mutation.

    PubMed

    Lanzicher, Thomas; Martinelli, Valentina; Long, Carlin S; Del Favero, Giorgia; Puzzi, Luca; Borelli, Massimo; Mestroni, Luisa; Taylor, Matthew R G; Sbaizero, Orfeo

    2015-01-01

    Previous investigations suggested that lamin A/C gene (LMNA) mutations, which cause a variety of human diseases including muscular dystrophies and cardiomyopathies, alter the nuclear mechanical properties. We hypothesized that biomechanical changes may extend beyond the nucleus.

  19. AFM single-cell force spectroscopy links altered nuclear and cytoskeletal mechanics to defective cell adhesion in cardiac myocytes with a nuclear lamin mutation

    PubMed Central

    Lanzicher, Thomas; Martinelli, Valentina; Long, Carlin S; Del Favero, Giorgia; Puzzi, Luca; Borelli, Massimo; Mestroni, Luisa; Taylor, Matthew R G; Sbaizero, Orfeo

    2015-01-01

    Previous investigations suggested that lamin A/C gene (LMNA) mutations, which cause a variety of human diseases including muscular dystrophies and cardiomyopathies, alter the nuclear mechanical properties. We hypothesized that biomechanical changes may extend beyond the nucleus. PMID:26309016

  20. Quantifying the effect of electric current on cell adhesion studied by single-cell force spectroscopy.

    PubMed

    Jaatinen, Leena; Young, Eleanore; Hyttinen, Jari; Vörös, János; Zambelli, Tomaso; Demkó, László

    2016-03-20

    This study presents the effect of external electric current on the cell adhesive and mechanical properties of the C2C12 mouse myoblast cell line. Changes in cell morphology, viability, cytoskeleton, and focal adhesion structure were studied by standard staining protocols, while single-cell force spectroscopy based on the fluidic force microscopy technology provided a rapid, serial quantification and detailed analysis of cell adhesion and its dynamics. The setup allowed measurements of adhesion forces up to the μN range, and total detachment distances over 40 μm. Force-distance curves have been fitted with a simple elastic model including a cell detachment protocol in order to estimate the Young's modulus of the cells, as well as to reveal changes in the dynamic properties as functions of the applied current dose. While the cell spreading area decreased monotonously with increasing current doses, small current doses resulted only in differences related to cell elasticity. Current doses above 11 As/m(2), however, initiated more drastic changes in cell morphology, viability, cellular structure, as well as in properties related to cell adhesion. The observed differences, eventually leading to cell death toward higher doses, might originate from both the decrease in pH and the generation of reactive oxygen species.

  1. Atomic force spectroscopy and density-functional study of graphene corrugation on Ru(0001)

    NASA Astrophysics Data System (ADS)

    Voloshina, Elena; Dedkov, Yuriy

    2016-06-01

    Graphene, the thinnest material in the world, can form moiré structures on different substrates, including graphite, h -BN, or metal surfaces. In such systems, the structure of graphene, i.e., its corrugation, as well as its electronic and elastic properties, are defined by the combination of the system geometry and local interaction strength at the interface. The corrugation in such structures on metals is heavily extracted from diffraction or local probe microscopy experiments, and it can be obtained only via comparison with theoretical data, which usually simulate the experimental findings. Here we show that graphene corrugation on metals can be measured directly employing atomic force spectroscopy, and the obtained value coincides with state-of-the-art theoretical results. The presented results demonstrate an unexpected space selectivity for the Δ f (z ) signal in the atomic force spectroscopy in the moiré graphene lattice on Ru(0001), which is explained by the different response of the graphene layer on the indentation process. We also address the elastic reaction of the formed graphene nanodoms on the indentation process by the scanning tip that is important for the modeling and fabrication of graphene-based nanoresonators on the nanoscale.

  2. Amyloid beta oligomers induce neuronal elasticity changes in age-dependent manner: a force spectroscopy study on living hippocampal neurons.

    PubMed

    Ungureanu, Andreea-Alexandra; Benilova, Iryna; Krylychkina, Olga; Braeken, Dries; De Strooper, Bart; Van Haesendonck, Chris; Dotti, Carlos G; Bartic, Carmen

    2016-01-01

    Small soluble species of amyloid-beta (Aβ) formed during early peptide aggregation stages are responsible for several neurotoxic mechanisms relevant to the pathology of Alzheimer's disease (AD), although their interaction with the neuronal membrane is not completely understood. This study quantifies the changes in the neuronal membrane elasticity induced by treatment with the two most common Aβ isoforms found in AD brains: Aβ40 and Aβ42. Using quantitative atomic force microscopy (AFM), we measured for the first time the static elastic modulus of living primary hippocampal neurons treated with pre-aggregated Aβ40 and Aβ42 soluble species. Our AFM results demonstrate changes in the elasticity of young, mature and aged neurons treated for a short time with the two Aβ species pre-aggregated for 2 hours. Neurons aging under stress conditions, showing aging hallmarks, are the most susceptible to amyloid binding and show the largest decrease in membrane stiffness upon Aβ treatment. Membrane stiffness defines the way in which cells respond to mechanical forces in their environment and has been shown to be important for processes such as gene expression, ion-channel gating and neurotransmitter vesicle transport. Thus, one can expect that changes in neuronal membrane elasticity might directly induce functional changes related to neurodegeneration.

  3. Amyloid beta oligomers induce neuronal elasticity changes in age-dependent manner: a force spectroscopy study on living hippocampal neurons

    PubMed Central

    Ungureanu, Andreea-Alexandra; Benilova, Iryna; Krylychkina, Olga; Braeken, Dries; De Strooper, Bart; Van Haesendonck, Chris; Dotti, Carlos G.; Bartic, Carmen

    2016-01-01

    Small soluble species of amyloid-beta (Aβ) formed during early peptide aggregation stages are responsible for several neurotoxic mechanisms relevant to the pathology of Alzheimer’s disease (AD), although their interaction with the neuronal membrane is not completely understood. This study quantifies the changes in the neuronal membrane elasticity induced by treatment with the two most common Aβ isoforms found in AD brains: Aβ40 and Aβ42. Using quantitative atomic force microscopy (AFM), we measured for the first time the static elastic modulus of living primary hippocampal neurons treated with pre-aggregated Aβ40 and Aβ42 soluble species. Our AFM results demonstrate changes in the elasticity of young, mature and aged neurons treated for a short time with the two Aβ species pre-aggregated for 2 hours. Neurons aging under stress conditions, showing aging hallmarks, are the most susceptible to amyloid binding and show the largest decrease in membrane stiffness upon Aβ treatment. Membrane stiffness defines the way in which cells respond to mechanical forces in their environment and has been shown to be important for processes such as gene expression, ion-channel gating and neurotransmitter vesicle transport. Thus, one can expect that changes in neuronal membrane elasticity might directly induce functional changes related to neurodegeneration. PMID:27173984

  4. Optimization of phase contrast in bimodal amplitude modulation AFM.

    PubMed

    Damircheli, Mehrnoosh; Payam, Amir F; Garcia, Ricardo

    2015-01-01

    Bimodal force microscopy has expanded the capabilities of atomic force microscopy (AFM) by providing high spatial resolution images, compositional contrast and quantitative mapping of material properties without compromising the data acquisition speed. In the first bimodal AFM configuration, an amplitude feedback loop keeps constant the amplitude of the first mode while the observables of the second mode have not feedback restrictions (bimodal AM). Here we study the conditions to enhance the compositional contrast in bimodal AM while imaging heterogeneous materials. The contrast has a maximum by decreasing the amplitude of the second mode. We demonstrate that the roles of the excited modes are asymmetric. The operational range of bimodal AM is maximized when the second mode is free to follow changes in the force. We also study the contrast in trimodal AFM by analyzing the kinetic energy ratios. The phase contrast improves by decreasing the energy of second mode relative to those of the first and third modes.

  5. Optimization of phase contrast in bimodal amplitude modulation AFM

    PubMed Central

    Damircheli, Mehrnoosh; Payam, Amir F

    2015-01-01

    Summary Bimodal force microscopy has expanded the capabilities of atomic force microscopy (AFM) by providing high spatial resolution images, compositional contrast and quantitative mapping of material properties without compromising the data acquisition speed. In the first bimodal AFM configuration, an amplitude feedback loop keeps constant the amplitude of the first mode while the observables of the second mode have not feedback restrictions (bimodal AM). Here we study the conditions to enhance the compositional contrast in bimodal AM while imaging heterogeneous materials. The contrast has a maximum by decreasing the amplitude of the second mode. We demonstrate that the roles of the excited modes are asymmetric. The operational range of bimodal AM is maximized when the second mode is free to follow changes in the force. We also study the contrast in trimodal AFM by analyzing the kinetic energy ratios. The phase contrast improves by decreasing the energy of second mode relative to those of the first and third modes. PMID:26114079

  6. The thermal near-field: Coherence, spectroscopy, heat-transfer, and optical forces

    NASA Astrophysics Data System (ADS)

    Jones, Andrew C.; O'Callahan, Brian T.; Yang, Honghua U.; Raschke, Markus B.

    2013-12-01

    One of the most universal physical processes shared by all matter at finite temperature is the emission of thermal radiation. The experimental characterization and theoretical description of far-field black-body radiation was a cornerstone in the development of modern physics with the groundbreaking contributions from Gustav Kirchhoff and Max Planck. With its origin in thermally driven fluctuations of the charge carriers, thermal radiation reflects the resonant and non-resonant dielectric properties of media, which is the basis for far-field thermal emission spectroscopy. However, associated with the underlying fluctuating optical source polarization are fundamentally distinct spectral, spatial, resonant, and coherence properties of the evanescent thermal near-field. These properties have been recently predicted theoretically and characterized experimentally for systems with thermally excited molecular, surface plasmon polariton (SPP), and surface phonon polariton (SPhP) resonances. We review, starting with the early historical developments, the emergence of theoretical models, and the description of the thermal near-field based on the fluctuation-dissipation theory and in terms of the electromagnetic local density of states (EM-LDOS). We discuss the optical and spectroscopic characterization of distance dependence, magnitude, spectral distribution, and coherence of evanescent thermal fields. Scattering scanning near-field microscopy proved instrumental as an enabling technique for the investigations of several of these fundamental thermal near-field properties. We then discuss the role of thermal fields in nano-scale heat transfer and optical forces, and the correlation to the van der Waals, Casimir, and Casimir-Polder forces. We conclude with an outlook on the possibility of intrinsic and extrinsic resonant manipulation of optical forces, control of nano-scale radiative heat transfer with optical antennas and metamaterials, and the use of thermal infrared near

  7. Thermo-magnetic behaviour of AFM-MFM cantilevers

    NASA Astrophysics Data System (ADS)

    Kumar, M.; Arinero, R.; Bergez, W.; Tordjeman, Ph

    2015-08-01

    Atomic force microscopy (AFM) experiments were performed to study the behaviour of AFM cantilevers under an external magnetic field B and temperature field produced by a coil with an iron core. Four cantilever types were studied. Forces were measured for different B values and at various coil-to-cantilever separation distances. The results were analysed on the basis of a phenomenological model. This model contains the contribution of two terms, one monopole-monopole interaction at short distance, and one apparent paramagnetic interaction in \

  8. Nanomechanics of new materials — AFM and computer modelling studies of trichoptera silk

    NASA Astrophysics Data System (ADS)

    Strzelecki, Janusz; Strzelecka, Joanna; Mikulska, Karolina; Tszydel, Mariusz; Balter, Aleksander; Nowak, Wiesław

    2011-04-01

    Caddisfly (Trichopera) can glue diverse material underwater with a silk fiber. This makes it a particularly interesting subject for biomimetcs. Better understanding of silk composition and structure could lead to an adhesive capable to close bleeding wounds or to new biomaterials. However, while spiderweb or silkworm secretion is well researched, caddisfly silk is still poorly understood. Here we report a first nanomechanical analysis of H. Angustipennis caddisfly silk fiber. An Atomic Force Microscope (AFM) imaging shows dense 150 nm bumps on silk surface, which can be identified as one of features responsible for its outstanding adhesive properties. AFM force spectroscopy at the fiber surface showed, among others, characteristic saw like pattern. This pattern is attributed to sacrificial bond stretching and enhances energy dissipation in mechanical deformation. Similarities of some force curves observed on Tegenaria domestica spiderweb and caddisfly silk are also discussed. Steered Molecular Dynamics simulations revealed that the strength of short components of Fib-H HA species molecules, abundant in Trichoptera silk is critically dependent on calcium presence.

  9. Supramolecular self-assembly of linear oligosilsesquioxanes on mica--AFM surface imaging and hydrophilicity studies.

    PubMed

    Kowalewska, Anna; Nowacka, Maria; Tracz, Adam; Makowski, Tomasz

    2015-06-28

    Linear oligomeric [2-(carboxymethylthio)ethylsilsesquioxanes] (LPSQ-COOH) adsorb spontaneously on muscovite mica and form smooth, well-ordered lamellar structures at the liquid-solid interface. Side carboxylic groups, having donor-acceptor character with regard to hydrogen bonds, are engaged both in multipoint molecule-to-substrate interactions and intermolecular cross-linking. The unique arrangement of silsesquioxane macromolecules, with COOH groups situated at the interface with air, produces highly hydrophilic surfaces of good thermal and solvolytic stability. Supramolecular assemblies of LPSQ-COOH were studied using atomic force microscopy (AFM), angle-resolved X-ray photoelectron spectroscopy (ARXPS) and attenuated total reflectance (ATR) FTIR spectroscopy. Comparative height profile analysis combined with ATR-FTIR studies of the spectral regions characteristic of carboxylic groups and C1s core level envelope by XPS confirmed specific interactions between LPSQ-COOH and mica.

  10. Multi-scale modeling of biophysical phenomena: ionic transport, biomineralization, and force spectroscopy

    NASA Astrophysics Data System (ADS)

    Kelly, Mark A.

    2011-07-01

    Biophysics is the study of the complex physical processes occurring in biological systems that are responsible for life. This dissertation addresses three important topics in biophysics: ionic transport, biomineralization, and force spectroscopy. Ionic transport involves the passage of ions through a special class of hollow, transmembrane proteins called ion channels which regulate the movement of charged species across nearly all biological membranes with varying degrees of specificity. Despite the fundamental importance of these channels to many physiological processes little is known about how channel structure and composition couple to determine its function. Deriving inspiration from these systems, a simple computational platform is developed to study the salient features of these channels in order to better understand the fundamental physics of these systems. The results of this work indicate that a converging-diverging region formed within the pore to create a single constriction is the most effective method to regulate the passage of ions through the pore. By controlling the geometry of the constriction the local potential and chemical gradients can be manipulated to tailor the channel for specific applications. The process of selective extraction and incorporation of local elements from the surrounding environment into functional structures under strict biological control is known as biomineralization. As an initial step to gain a more fundamental understanding of directed crystallization of zinc oxide molecular dynamics simulations were performed to study the conformational behavior of two experimentally derived biomimetic peptides in a precursor solution. Substantial differences in the conformational properties and affinity for zinc and hydroxide ions in solution were observed. These findings are in qualitative agreement with experimental observations. The mechanical response of biopolymers such as RNA and DNA to externally applied forces is a topic that

  11. Decoupling indirect topographic cross-talk in band excitation piezoresponse force microscopy imaging and spectroscopy

    NASA Astrophysics Data System (ADS)

    Yang, Sang Mo; Mazet, Lucie; Okatan, M. Baris; Jesse, Stephen; Niu, Gang; Schroeder, Thomas; Schamm-Chardon, Sylvie; Dubourdieu, Catherine; Baddorf, Arthur P.; Kalinin, Sergei V.

    2016-06-01

    All scanning probe microscopies are subjected to topographic cross-talk, meaning the topography-related contrast in functional images. Here, we investigate the signatures of indirect topographic cross-talk in piezoresponse force microscopy (PFM) imaging and spectroscopy and its decoupling using band excitation (BE) method in ferroelectric BaTiO3 deposited on the Si substrates with free standing nanopillars of diameter 50 nm. Comparison between the single-frequency PFM and BE-PFM results shows that the measured signal can be significantly distorted by topography-induced shifts in the contact resonance frequency and cantilever transfer function. However, with proper correction, such shifts do not affect PFM imaging and hysteresis loop measurements. This suggests the necessity of an advanced approach, such as BE-PFM, for detection of intrinsic sample piezoresponse on the topographically non-uniform surfaces.

  12. Decoupling indirect topographic cross-talk in band excitation piezoresponse force microscopy imaging and spectroscopy

    DOE PAGES

    Mazet, Lucie; Jesse, Stephen; Niu, Gang; Schroeder, Thomas; Schamm-Chardon, Sylvie; Dubourdieu, Catherine; Baddorf, Arthur P.; Kalinin, Sergei V.; Yang, Sang Mo; Okatan, M. Baris

    2016-06-20

    Here, all scanning probe microscopies are subjected to topographic cross-talk, meaning the topography-related contrast in functional images. Here, we investigate the signatures of indirect topographic cross-talk in piezoresponse force microscopy (PFM) imaging and spectroscopy and its decoupling using band excitation (BE) method in ferroelectric BaTiO3 deposited on the Si substrates with free standing nanopillars of diameter 50 nm. Comparison between the single-frequency PFM and BE-PFM results shows that the measured signal can be significantly distorted by topography-induced shifts in the contact resonance frequency and cantilever transfer function. However, with proper correction, such shifts do not affect PFM imaging and hysteresismore » loop measurements. This suggests the necessity of an advanced approach, such as BE-PFM, for detection of intrinsic sample piezoresponse on the topographically non-uniform surfaces.« less

  13. Thermal activation at moderate-to-high and high damping: Finite barrier effects and force spectroscopy

    NASA Astrophysics Data System (ADS)

    Mazo, J. J.; Fajardo, O. Y.; Zueco, D.

    2013-03-01

    We study the thermal escape problem in the moderate-to-high and high damping regime of a system with a parabolic barrier. We present a formula that matches our numerical results accounting for finite barrier effects, and compare it with previous works. We also show results for the full damping range. We quantitatively study some aspects on the relation between mean first passage time and the definition of an escape rate. To finish, we apply our results and considerations in the framework of force spectroscopy problems. We study the differences on the predictions using the different theories and discuss the role of γ dot{F} as the relevant parameter at high damping.

  14. In Situ Adsorption Studies at the Solid/Liquid Interface:Characterization of Biological Surfaces and Interfaces Using SumFrequency Generation Vibrational Spectroscopy, Atomic Force Microscopy,and Quartz Crystal Microbalance

    SciTech Connect

    Phillips, Diana Christine

    2006-01-01

    Sum frequency generation (SFG) vibrational spectroscopy, atomic force microscopy (AFM), and quartz crystal microbalance (QCM) have been used to study the molecular surface structure, surface topography and mechanical properties, and quantitative adsorbed amount of biological molecules at the solid-liquid interface. The molecular-level behavior of designed peptides adsorbed on hydrophobic polystyrene and hydrophilic silica substrates has been examined as a model of protein adsorption on polymeric biomaterial surfaces. Proteins are such large and complex molecules that it is difficult to identify the features in their structure that lead to adsorption and interaction with solid surfaces. Designed peptides which possess secondary structure provide simple model systems for understanding protein adsorption. Depending on the amino acid sequence of a peptide, different secondary structures (α-helix and β-sheet) can be induced at apolar (air/liquid or air/solid) interfaces. Having a well-defined secondary structure allows experiments to be carried out under controlled conditions, where it is possible to investigate the affects of peptide amino acid sequence and chain length, concentration, buffering effects, etc. on adsorbed peptide structure. The experiments presented in this dissertation demonstrate that SFG vibrational spectroscopy can be used to directly probe the interaction of adsorbing biomolecules with a surface or interface. The use of well designed model systems aided in isolation of the SFG signal of the adsorbing species, and showed that surface functional groups of the substrate are sensitive to surface adsorbates. The complementary techniques of AFM and QCM allowed for deconvolution of the effects of surface topography and coverage from the observed SFG spectra. Initial studies of biologically relevant surfaces are also presented: SFG spectroscopy was used to study the surface composition of common soil bacteria for use in bioremediation of nuclear waste.

  15. Atomic Force Microscope Mediated Chromatography

    NASA Technical Reports Server (NTRS)

    Anderson, Mark S.

    2013-01-01

    The atomic force microscope (AFM) is used to inject a sample, provide shear-driven liquid flow over a functionalized substrate, and detect separated components. This is demonstrated using lipophilic dyes and normal phase chromatography. A significant reduction in both size and separation time scales is achieved with a 25-micron-length column scale, and one-second separation times. The approach has general applications to trace chemical and microfluidic analysis. The AFM is now a common tool for ultra-microscopy and nanotechnology. It has also been demonstrated to provide a number of microfluidic functions necessary for miniaturized chromatography. These include injection of sub-femtoliter samples, fluidic switching, and sheardriven pumping. The AFM probe tip can be used to selectively remove surface layers for subsequent microchemical analysis using infrared and tip-enhanced Raman spectroscopy. With its ability to image individual atoms, the AFM is a remarkably sensitive detector that can be used to detect separated components. These diverse functional components of microfluidic manipulation have been combined in this work to demonstrate AFM mediated chromatography. AFM mediated chromatography uses channel-less, shear-driven pumping. This is demonstrated with a thin, aluminum oxide substrate and a non-polar solvent system to separate a mixture of lipophilic dyes. In conventional chromatographic terms, this is analogous to thin-layer chromatography using normal phase alumina substrate with sheardriven pumping provided by the AFM tip-cantilever mechanism. The AFM detection of separated components is accomplished by exploiting the variation in the localized friction of the separated components. The AFM tip-cantilever provides the mechanism for producing shear-induced flows and rapid pumping. Shear-driven chromatography (SDC) is a relatively new concept that overcomes the speed and miniaturization limitations of conventional liquid chromatography. SDC is based on a

  16. Conductive-probe AFM characterization of graphene sheets bonded to gold surfaces

    NASA Astrophysics Data System (ADS)

    Hauquier, Fanny; Alamarguy, David; Viel, Pascal; Noël, Sophie; Filoramo, Arianna; Huc, Vincent; Houzé, Frédéric; Palacin, Serge

    2012-01-01

    Conducting probe atomic force microscopy (CP-AFM) has been used to perform mechanical and electrical experiments on graphene layers bonded to polyaminophenylene (PAP) films grafted on gold substrates. This technique is a new approach for the characterization of graphene sheets and represents a complementary tool to Raman spectroscopy. The combination of friction and electrical imaging reveals that different stacked graphene sheets have been successfully distinguished from each other and from the underlying PAP films. Lateral force microscopy has shown that the friction is greatly reduced on graphene sheets in comparison with the organic coating. The electrical resistance images show very different local conduction properties which can be linked to the number of underlying graphene sheets. The resistance decreases very slowly when the normal load increases. Current-voltage curves display characteristics of metal-molecule-metal junctions.

  17. Nanoscopic oxidation of p-type and un-doped Si (100) surfaces using un-externally biased atomic force microscope tips (AFM) in the presence of selected organic solvents

    NASA Astrophysics Data System (ADS)

    McCausland, Jeffrey; Withanage, Sajeevi; Mallik, Robert; Lyuksyutov, Sergei

    A conductive un-biased AFM tip oscillating above p-type or un-doped Si (100) treated with toluene, butan-2-ol, and propan-2-ol creates nanostructures ranging in height from 1-100 nm. The tip was oscillated in ambient conditions (30-70% Rel. Humidity) at frequencies in the 102 kHz range. It was repeatable with various concentrations of solvent in aqueous solution. It is suggested that mechanical oscillations of the AFM tip polarizes the solvent molecules deposited on the surface resulting in electron transfer from the tip to the surface followed by feature formation. This process effectively creates an electrochemical cell at the microscopic level and the miscibility of the solvents is the key to enabling the process. Species which ionize during the process may be consumed in irreversible reactions whereas the alcohols act as catalysts and are not consumed. The influence of boron defects in the Si substrates is also discussed. It appears that the observed oxidation is different from all other similar reported phenomena including local anodic oxidation, and chemo-mechanical lithographic techniques utilizing AFM.

  18. Characterization of the molecular structure and mechanical properties of polymer surfaces and protein/polymer interfaces by sum frequency generation vibrational spectroscopy and atomic force microscopy

    SciTech Connect

    Koffas, Telly Stelianos

    2004-01-01

    Sum frequency generation (SFG) vibrational spectroscopy, atomic force microscopy (AFM), and other complementary surface-sensitive techniques have been used to study the surface molecular structure and surface mechanical behavior of biologically-relevant polymer systems. SFG and AFM have emerged as powerful analytical tools to deduce structure/property relationships, in situ, for polymers at air, liquid and solid interfaces. The experiments described in this dissertation have been performed to understand how polymer surface properties are linked to polymer bulk composition, substrate hydrophobicity, changes in the ambient environment (e.g., humidity and temperature), or the adsorption of macromolecules. The correlation of spectroscopic and mechanical data by SFG and AFM can become a powerful methodology to study and engineer materials with tailored surface properties. The overarching theme of this research is the interrogation of systems of increasing structural complexity, which allows us to extend conclusions made on simpler model systems. We begin by systematically describing the surface molecular composition and mechanical properties of polymers, copolymers, and blends having simple linear architectures. Subsequent chapters focus on networked hydrogel materials used as soft contact lenses and the adsorption of protein and surfactant at the polymer/liquid interface. The power of SFG is immediately demonstrated in experiments which identify the chemical parameters that influence the molecular composition and ordering of a polymer chain's side groups at the polymer/air and polymer/liquid interfaces. In general, side groups with increasingly greater hydrophobic character will be more surface active in air. Larger side groups impose steric restrictions, thus they will tend to be more randomly ordered than smaller hydrophobic groups. If exposed to a hydrophilic environment, such as water, the polymer chain will attempt to orient more of its hydrophilic groups to the

  19. Examination of dentin surface using AFM (our experience).

    PubMed

    Zapletalová, Zdenka; Kubínek, Roman; Vůjtek, Milan; Novotný, Radko

    2004-01-01

    Atomic force microscopy (AFM) as one the technique of Scanning Probe Microscopy is useful for imaging of surface structure. This method can yield three-dimensional high-resolution topographic images of sample surfaces by using a scanning technique for conductors and insulators on atomic scale. It is based upon mapping of atomic-forces on a surface of an investigated sample. The method is useful not only in physics and chemistry; it can be also applied in biological fields. Special construction of AFM scanner enables to follow biological samples in liquid environments. Artifacts caused by dehydration of samples are removed this way. Dentin of human teeth is a vital hydrated tissue. It is strongly sensitive to dehydration and drying that are commonly used in preparation of samples in examinations by Scanning Electron Microscopy (SEM). We describe our experience in examination of dentin surfaces of extracted human third molars using contact method of AFM under moist conditions.

  20. Direct Force Measurements of Receptor-Ligand Interactions on Living Cells

    NASA Astrophysics Data System (ADS)

    Eibl, Robert H.

    The characterization of cell adhesion between two living cells at the level of single receptor-ligand bonds is an experimental challenge. This chapter describes how the extremely sensitive method of atomic force microscopy (AFM) based force spectroscopy can be applied to living cells in order to probe for cell-to-cell or cell-to-substrate interactions mediated by single pairs of adhesion receptors. In addition, it is outlined how single-molecule AFM force spectroscopy can be used to detect physiologic changes of an adhesion receptor in a living cell. This force spectroscopy allows us to detect in living cells rapidly changing, chemokine SDF-1 triggered activation states of single VLA-4 receptors. This recently developed AFM application will allow for the detailed investigation of the integrin-chemokine crosstalk of integrin activation mechanisms and on how other adhesion receptors are modulated in health and disease. As adhesion molecules, living cells and even bacteria can be studied by single-molecule AFM force spectroscopy, this method is set to become a powerful tool that can not only be used in biophysics, but in cell biology as well as in immunology and cancer research.

  1. Stability of bacteriorhodopsin alpha-helices and loops analyzed by single-molecule force spectroscopy.

    PubMed Central

    Müller, Daniel J; Kessler, Max; Oesterhelt, Filipp; Möller, Clemens; Oesterhelt, Dieter; Gaub, Hermann

    2002-01-01

    The combination of high-resolution atomic force microscopy imaging and single-molecule force spectroscopy allows the identification, selection, and mechanical investigation of individual proteins. In a recent paper we had used this technique to unfold and extract single bacteriorhodopsins (BRs) from native purple membrane patches. We show that subsets of the unfolding spectra can be classified and grouped to reveal detailed insight into the individualism of the unfolding pathways. We have further developed this technique and analysis to report here on the influence of pH effects and local mutations on the stability of individual structural elements of BR against mechanical unfolding. We found that, although the seven transmembrane alpha-helices predominantly unfold in pairs, each of the helices may also unfold individually and in some cases even only partially. Additionally, intermittent states in the unfolding process were found, which are associated with the stretching of the extracellular loops connecting the alpha-helices. This suggests that polypeptide loops potentially act as a barrier to unfolding and contribute significantly to the structural stability of BR. Chemical removal of the Schiff base, the covalent linkage of the photoactive retinal to the helix G, resulted in a predominantly two-step unfolding of this helix. It is concluded that the covalent linkage of the retinal to helix G stabilizes the structure of BR. Trapping mutant D96N in the M state of the proton pumping photocycle did not affect the unfolding barriers of BR. PMID:12496125

  2. Unraveling the complexity of the interactions of DNA nucleotides with gold by single molecule force spectroscopy.

    PubMed

    Bano, Fouzia; Sluysmans, Damien; Wislez, Arnaud; Duwez, Anne-Sophie

    2015-12-14

    Addressing the effect of different environmental factors on the adsorption of DNA to solid supports is critical for the development of robust miniaturized devices for applications ranging from biosensors to next generation molecular technology. Most of the time, thiol-based chemistry is used to anchor DNA on gold - a substrate commonly used in nanotechnology - and little is known about the direct interaction between DNA and gold. So far there have been no systematic studies on the direct adsorption behavior of the deoxyribonucleotides (i.e., a nitrogenous base, a deoxyribose sugar, and a phosphate group) and on the factors that govern the DNA-gold bond strength. Here, using single molecule force spectroscopy, we investigated the interaction of the four individual nucleotides, adenine, guanine, cytosine, and thymine, with gold. Experiments were performed in three salinity conditions and two surface dwell times to reveal the factors that influence nucleotide-Au bond strength. Force data show that, at physiological ionic strength, adenine-Au interactions are stronger, asymmetrical and independent of surface dwell time as compared to cytosine-Au and guanine-Au interactions. We suggest that in these conditions only adenine is able to chemisorb on gold. A decrease of the ionic strength significantly increases the bond strength for all nucleotides. We show that moderate ionic strength along with longer surface dwell period suggest weak chemisorption also for cytosine and guanine.

  3. Integrin and glycocalyx mediated contributions to cell adhesion identified by single cell force spectroscopy

    NASA Astrophysics Data System (ADS)

    Boettiger, D.; Wehrle-Haller, B.

    2010-05-01

    The measurement of cell adhesion using single cell force spectroscopy methods was compared with earlier methods for measuring cell adhesion. This comparison provided a means and rationale for separating components of the measurement retract curve that were due to interactions between the substrate and the glycocalyx, and interactions that were due to cell surface integrins binding to a substrate-bound ligand. The glycocalyx adhesion was characterized by multiple jumps with dispersed jump sizes that extended from 5 to 30 µm from the origin. The integrin mediated adhesion was represented by the Fmax (maximum detachment force), was generally within the first 5 µm and commonly detached with a single rupture cascade. The integrin peak (Fmax) increases with time and the rate of increase shows large cell to cell variability with a peak ~ 50 nN s - 1 and an average rate of increase of 75 pN s - 1. This is a measure of the rate of increase in the number of adhesive integrin-ligand bonds/cell as a function of contact time.

  4. Unraveling the complexity of the interactions of DNA nucleotides with gold by single molecule force spectroscopy

    NASA Astrophysics Data System (ADS)

    Bano, Fouzia; Sluysmans, Damien; Wislez, Arnaud; Duwez, Anne-Sophie

    2015-11-01

    Addressing the effect of different environmental factors on the adsorption of DNA to solid supports is critical for the development of robust miniaturized devices for applications ranging from biosensors to next generation molecular technology. Most of the time, thiol-based chemistry is used to anchor DNA on gold - a substrate commonly used in nanotechnology - and little is known about the direct interaction between DNA and gold. So far there have been no systematic studies on the direct adsorption behavior of the deoxyribonucleotides (i.e., a nitrogenous base, a deoxyribose sugar, and a phosphate group) and on the factors that govern the DNA-gold bond strength. Here, using single molecule force spectroscopy, we investigated the interaction of the four individual nucleotides, adenine, guanine, cytosine, and thymine, with gold. Experiments were performed in three salinity conditions and two surface dwell times to reveal the factors that influence nucleotide-Au bond strength. Force data show that, at physiological ionic strength, adenine-Au interactions are stronger, asymmetrical and independent of surface dwell time as compared to cytosine-Au and guanine-Au interactions. We suggest that in these conditions only adenine is able to chemisorb on gold. A decrease of the ionic strength significantly increases the bond strength for all nucleotides. We show that moderate ionic strength along with longer surface dwell period suggest weak chemisorption also for cytosine and guanine.Addressing the effect of different environmental factors on the adsorption of DNA to solid supports is critical for the development of robust miniaturized devices for applications ranging from biosensors to next generation molecular technology. Most of the time, thiol-based chemistry is used to anchor DNA on gold - a substrate commonly used in nanotechnology - and little is known about the direct interaction between DNA and gold. So far there have been no systematic studies on the direct

  5. Ultrafast optical pump-probe spectroscopy is used to reveal the coexistence of coupled antiferromagnetic (AFM)/ferroelectric (FE) and ferromagnetic (FM) orders in multiferroic TbMnO3 films, which can guide researchers in creating new kinds of multiferroic materials.

    SciTech Connect

    Qi, Jingbo; Zhu, Jianxin; Trugman, Stuart A.; Taylor, Antoinette; Jia, Quanxi; Prasankumar, Rohit

    2012-07-06

    , experimental techniques capable of dynamically unraveling the interplay between these degrees of freedom on an ultrafast timescale are needed. Here, we use ultrafast optical pump-probe spectroscopy to reveal coexisting coupled magnetic orders in epitaxial TMO thin films grown on (001)-STO, which were not observed in previous work. Our temperature (T)-dependent transient differential reflectivity ({Delta}R/R) measurements show clear signatures of sinusoidal AFM, spiral AFM (FE) and FM phases developing as the film thickness changes. We carry out first-principle density functional theory (DFT) calculations to explain the coupling between AFM/FE and FM orders. These results reveal that the coupling between different magnetic orders observed in our multiferroic TMO thin films may offer greater control of functionality as compared to bulk single crystal multiferroics.

  6. Can Dissipative Properties of Single Molecules Be Extracted from a Force Spectroscopy Experiment?

    PubMed

    Benedetti, Fabrizio; Gazizova, Yulia; Kulik, Andrzej J; Marszalek, Piotr E; Klinov, Dmitry V; Dietler, Giovanni; Sekatskii, Sergey K

    2016-09-20

    We performed dynamic force spectroscopy of single dextran and titin I27 molecules using small-amplitude and low-frequency (40-240 Hz) dithering of an atomic force microscope tip excited by a sine wave voltage fed onto the tip-carrying piezo. We show that for such low-frequency dithering experiments, recorded phase information can be unambiguously interpreted within the framework of a transparent theoretical model that starts from a well-known partial differential equation to describe the dithering of an atomic force microscope cantilever and a single molecule attached to its end system, uses an appropriate set of initial and boundary conditions, and does not exploit any implicit suggestions. We conclude that the observed phase (dissipation) signal is due completely to the dissipation related to the dithering of the cantilever itself (i.e., to the change of boundary conditions in the course of stretching). For both cases, only the upper bound of the dissipation of a single molecule has been established as not exceeding 3⋅10(-7)kg/s. We compare our results with previously reported measurements of the viscoelastic properties of single molecules, and we emphasize that extreme caution must be taken in distinguishing between the dissipation related to the stretched molecule and the dissipation that originates from the viscous damping of the dithered cantilever. We also present the results of an amplitude channel data analysis, which reveal that the typical values of the spring constant of a I27 molecule at the moment of module unfolding are equal to 4±1.5mN/m, and the typical values of the spring constant of dextran at the moment of chair-boat transition are equal to 30-50mN/m. PMID:27653475

  7. Can Dissipative Properties of Single Molecules Be Extracted from a Force Spectroscopy Experiment?

    PubMed

    Benedetti, Fabrizio; Gazizova, Yulia; Kulik, Andrzej J; Marszalek, Piotr E; Klinov, Dmitry V; Dietler, Giovanni; Sekatskii, Sergey K

    2016-09-20

    We performed dynamic force spectroscopy of single dextran and titin I27 molecules using small-amplitude and low-frequency (40-240 Hz) dithering of an atomic force microscope tip excited by a sine wave voltage fed onto the tip-carrying piezo. We show that for such low-frequency dithering experiments, recorded phase information can be unambiguously interpreted within the framework of a transparent theoretical model that starts from a well-known partial differential equation to describe the dithering of an atomic force microscope cantilever and a single molecule attached to its end system, uses an appropriate set of initial and boundary conditions, and does not exploit any implicit suggestions. We conclude that the observed phase (dissipation) signal is due completely to the dissipation related to the dithering of the cantilever itself (i.e., to the change of boundary conditions in the course of stretching). For both cases, only the upper bound of the dissipation of a single molecule has been established as not exceeding 3⋅10(-7)kg/s. We compare our results with previously reported measurements of the viscoelastic properties of single molecules, and we emphasize that extreme caution must be taken in distinguishing between the dissipation related to the stretched molecule and the dissipation that originates from the viscous damping of the dithered cantilever. We also present the results of an amplitude channel data analysis, which reveal that the typical values of the spring constant of a I27 molecule at the moment of module unfolding are equal to 4±1.5mN/m, and the typical values of the spring constant of dextran at the moment of chair-boat transition are equal to 30-50mN/m.

  8. Force.

    ERIC Educational Resources Information Center

    Gamble, Reed

    1989-01-01

    Discusses pupil misconceptions concerning forces. Summarizes some of Assessment of Performance Unit's findings on meaning of (1) force, (2) force and motion in one dimension and two dimensions, and (3) Newton's second law. (YP)

  9. Nano-Wilhelmy investigation of dynamic wetting properties of AFM tips through tip-nanobubble interaction

    NASA Astrophysics Data System (ADS)

    Wang, Yuliang; Wang, Huimin; Bi, Shusheng; Guo, Bin

    2016-07-01

    The dynamic wetting properties of atomic force microscopy (AFM) tips are of much concern in many AFM-related measurement, fabrication, and manipulation applications. In this study, the wetting properties of silicon and silicon nitride AFM tips are investigated through dynamic contact angle measurement using a nano-Wilhelmy balance based method. This is done by capillary force measurement during extension and retraction motion of AFM tips relative to interfacial nanobubbles. The working principle of the proposed method and mathematic models for dynamic contact angle measurement are presented. Geometric models of AFM tips were constructed using scanning electronic microscopy (SEM) images taken from different view directions. The detailed process of tip-nanobubble interaction was investigated using force-distance curves of AFM on nanobubbles. Several parameters including nanobubble height, adhesion and capillary force between tip and nanobubbles are extracted. The variation of these parameters was studied over nanobubble surfaces. The dynamic contact angles of the AFM tips were calculated from the capillary force measurements. The proposed method provides direct measurement of dynamic contact angles for AFM tips and can also be taken as a general approach for nanoscale dynamic wetting property investigation.

  10. Nano-Wilhelmy investigation of dynamic wetting properties of AFM tips through tip-nanobubble interaction

    PubMed Central

    Wang, Yuliang; Wang, Huimin; Bi, Shusheng; Guo, Bin

    2016-01-01

    The dynamic wetting properties of atomic force microscopy (AFM) tips are of much concern in many AFM-related measurement, fabrication, and manipulation applications. In this study, the wetting properties of silicon and silicon nitride AFM tips are investigated through dynamic contact angle measurement using a nano-Wilhelmy balance based method. This is done by capillary force measurement during extension and retraction motion of AFM tips relative to interfacial nanobubbles. The working principle of the proposed method and mathematic models for dynamic contact angle measurement are presented. Geometric models of AFM tips were constructed using scanning electronic microscopy (SEM) images taken from different view directions. The detailed process of tip-nanobubble interaction was investigated using force-distance curves of AFM on nanobubbles. Several parameters including nanobubble height, adhesion and capillary force between tip and nanobubbles are extracted. The variation of these parameters was studied over nanobubble surfaces. The dynamic contact angles of the AFM tips were calculated from the capillary force measurements. The proposed method provides direct measurement of dynamic contact angles for AFM tips and can also be taken as a general approach for nanoscale dynamic wetting property investigation. PMID:27452115

  11. Atomic force microscopy of Precambrian microscopic fossils.

    PubMed

    Kempe, André; Schopf, J William; Altermann, Wladyslaw; Kudryavtsev, Anatoliy B; Heckl, Wolfgang M

    2002-07-01

    Atomic force microscopy (AFM) is a technique used routinely in material science to image substances at a submicron (including nm) scale. We apply this technique to analysis of the fine structure of organic-walled Precambrian fossils, microscopic sphaeromorph acritarchs (cysts of planktonic unicellular protists) permineralized in approximately 650-million-year-old cherts of the Chichkan Formation of southern Kazakhstan. AFM images, backed by laser-Raman spectroscopic analysis of individual specimens, demonstrate that the walls of these petrified fossils are composed of stacked arrays of approximately 200-nm-sized angular platelets of polycyclic aromatic kerogen. Together, AFM and laser-Raman spectroscopy provide means by which to elucidate the submicron-scale structure of individual microscopic fossils, investigate the geochemical maturation of ancient organic matter, and, potentially, distinguish true fossils from pseudofossils and probe the mechanisms of fossil preservation by silica permineralization.

  12. Method and system for near-field spectroscopy using targeted deposition of nanoparticles

    NASA Technical Reports Server (NTRS)

    Anderson, Mark S. (Inventor)

    2012-01-01

    There is provided in one embodiment of the invention a method for analyzing a sample material using surface enhanced spectroscopy. The method comprises the steps of imaging the sample material with an atomic force microscope (AFM) to select an area of interest for analysis, depositing nanoparticles onto the area of interest with an AFM tip, illuminating the deposited nanoparticles with a spectrometer excitation beam, and disengaging the AFM tip and acquiring a localized surface enhanced spectrum. The method may further comprise the step of using the AFM tip to modulate the spectrometer excitation beam above the deposited nanoparticles to obtain improved sensitivity data and higher spatial resolution data from the sample material. The invention further comprises in one embodiment a system for analyzing a sample material using surface enhanced spectroscopy.

  13. AFM investigation of Martian soil simulants on micromachined Si substrates.

    PubMed

    Vijendran, S; Sykulska, H; Pike, W T

    2007-09-01

    The micro and nanostructures of Martian soil simulants with particles in the micrometre-size range have been studied using a combination of optical and atomic force microscopy (AFM) in preparation for the 2007 NASA Phoenix Mars Lander mission. The operation of an atomic force microscope on samples of micrometre-sized soil particles is a poorly investigated area where the unwanted interaction between the scanning tip and loose particles results in poor image quality and tip contamination by the sample. In order to mitigate these effects, etched silicon substrates with a variety of features have been used to facilitate the sorting and gripping of particles. From these experiments, a number of patterns were identified that were particularly good at isolating and immobilizing particles for AFM imaging. This data was used to guide the design of micromachined substrates for the Phoenix AFM. Both individual particles as well as aggregates were successfully imaged, and information on sizes, shapes and surface morphologies were obtained. This study highlights both the strengths and weaknesses of AFM for the potential in situ investigation of Martian soil and dust. Also presented are more general findings of the limiting operational constraints that exist when attempting the AFM of high aspect ratio particles with current technology. The performance of the final designs of the substrates incorporated on Phoenix will be described in a later paper. PMID:17760618

  14. AFM Structural Characterization of Drinking Water Biofilm under Physiological Conditions

    EPA Science Inventory

    Due to the complexity of mixed culture drinking water biofilm, direct visual observation under in situ conditions has been challenging. In this study, atomic force microscopy (AFM) revealed the three dimensional morphology and arrangement of drinking water relevant biofilm in air...

  15. Structure, cell wall elasticity and polysaccharide properties of living yeast cells, as probed by AFM

    NASA Astrophysics Data System (ADS)

    Alsteens, David; Dupres, Vincent; McEvoy, Kevin; Wildling, Linda; Gruber, Hermann J.; Dufrêne, Yves F.

    2008-09-01

    Although the chemical composition of yeast cell walls is known, the organization, assembly, and interactions of the various macromolecules remain poorly understood. Here, we used in situ atomic force microscopy (AFM) in three different modes to probe the ultrastructure, cell wall elasticity and polymer properties of two brewing yeast strains, i.e. Saccharomyces carlsbergensis and S. cerevisiae. Topographic images of the two strains revealed smooth and homogeneous cell surfaces, and the presence of circular bud scars on dividing cells. Nanomechanical measurements demonstrated that the cell wall elasticity of S. carlsbergensis is homogeneous. By contrast, the bud scar of S. cerevisiae was found to be stiffer than the cell wall, presumably due to the accumulation of chitin. Notably, single molecule force spectroscopy with lectin-modified tips revealed major differences in polysaccharide properties of the two strains. Polysaccharides were clearly more extended on S. cerevisiae, suggesting that not only oligosaccharides, but also polypeptide chains of the mannoproteins were stretched. Consistent with earlier cell surface analyses, these findings may explain the very different aggregation properties of the two organisms. This study demonstrates the power of using multiple complementary AFM modalities for probing the organization and interactions of the various macromolecules of microbial cell walls.

  16. Measuring cell wall elasticity on enteroaggregative Escherichia coli wild type and dispersin mutant by AFM

    SciTech Connect

    Beckmann, Melissa; Venkataraman, Sankar; Doktycz, Mitchel John; Nataro, James P; Sullivan, Claretta J; Morrell-Falvey, Jennifer L; Allison, David P

    2006-07-01

    Enteroaggregative Escherichia coli (EAEC) is pathogenic and produces severe diarrhea in humans. A mutant of EAEC that does not produce dispersin, a cell surface protein, is not pathogenic. It has been proposed that dispersin imparts a positive charge to the bacterial cell surface allowing the bacteria to colonize on the negatively charged intestinal mucosa. However, physical properties of the bacterial cell surface, such as rigidity, may be influenced by the presence of dispersin and may contribute to pathogenicity. Using the system developed in our laboratory for mounting and imaging bacterial cells by atomic force microscopy (AFM), in liquid, on gelatin coated mica surfaces, studies were initiated to measure cell surface elasticity. This was carried out in both wild type EAEC, that produces dispersin, and the mutant that does not produce dispersin. This was accomplished using AFM force-distance (FD) spectroscopy on the wild type and mutant grown in liquid or on solid medium. Images in liquid and in air of both the wild-type and mutant grown in liquid and on solid media are presented. This work represents an initial step in efforts to understand the pathogenic role of the dispersin protein in the wild-type bacteria.

  17. AFM Observation of Self-Assembled Monolayer Films on GaAs (110)

    NASA Astrophysics Data System (ADS)

    Ohno, Hirotaka; Motomatsu, Makoto; Mizutani, Wataru; Tokumoto, Hiroshi

    1995-02-01

    We have confirmed that a self-assembled monolayer (SAM) film of octadecanethiol (ODT), CH3(CH2)17SH, can be formed on a cleaved GaAs (110) surface, by using an atomic force microscope (AFM) and X-ray photoelectron spectroscopy (XPS). Circular depressions were observed on the surface after film formation. The area of the circular depressions increased with immersion time, indicating that the solution oxidized the interface between ODT molecules and the GaAs surface, resulting in removal of ODT molecules. The oxidation was considerably faster in pure ethanol solution than that in ODT solution, demonstrating that the SAM film protects the GaAs surface from oxidation. High-resolution lateral force microscope (LFM) images revealed a periodic structure that had two types of lines: periodic lines 0.57 nm apart and lines rotated 55° with respect to them. A structural model of the SAM successfully explained both the features in high-resolution LFM images and the depression depth observed in AFM images.

  18. The formation of liquid bridge in different operating modes of AFM

    NASA Astrophysics Data System (ADS)

    Wei, Zheng; Sun, Yan; Ding, WenXuan; Wang, ZaiRan

    2016-09-01

    The liquid bridge is one of the principal factors that cause artifacts in ambient-pressure atomic force microscope (AFM) images. Additionally, it is the main component of the adhesion force in ambient conditions. To understand the AFM imaging mechanism and the sample characteristics, it is essential to study the liquid bridge. This study interprets the physical mechanism involved in liquid bridge formation, which is composed of three different physical processes: the squeezing process, capillary condensation, and liquid film flow. We discuss the contributions of these three mechanisms to the volume and the capillary force of the liquid bridge in different AFM operation modes.

  19. Beyond topography - enhanced imaging of cometary dust with the MIDAS AFM

    NASA Astrophysics Data System (ADS)

    Bentley, M. S.; Torkar, K.; Jeszenszky, H.; Romstedt, J.

    2013-09-01

    The MIDAS atomic force microscope (AFM) onboard the Rosetta spacecraft is primarily designed to return the 3D shape and structure of cometary dust particles collected at comet 67P/Churyumov-Gerasimenko [1]. Commercial AFMs have, however, been further developed to measure many other sample properties. The possibilities to make such measurements with MIDAS are explored here.

  20. Mapping individual cosmid DNAs by direct AFM imaging.

    PubMed

    Allison, D P; Kerper, P S; Doktycz, M J; Thundat, T; Modrich, P; Larimer, F W; Johnson, D K; Hoyt, P R; Mucenski, M L; Warmack, R J

    1997-05-01

    Individual cosmid clones have been restriction mapped by directly imaging, with the atomic force microscope (AFM), a mutant EcoRI endonuclease site-specifically bound to DNA. Images and data are presented that locate six restriction sites, predicted from gel electrophoresis, on a 35-kb cosmid isolated from mouse chromosome 7. Measured distances between endonuclease molecules bound to lambda DNA, when compared to known values, demonstrate the accuracy of AFM mapping to better than 1%. These results may be extended to identify other important site-specific protein-DNA interactions, such as transcription factor and mismatch repair enzyme binding, difficult to resolve by current techniques.

  1. Cells mechanics with AFM: problems and solutions

    NASA Astrophysics Data System (ADS)

    Sokolov, Igor; Dokukin, Maxim; Guz, Nataliia

    2012-02-01

    Atomic force microscopy (AFM) is used to study mechanics of cells. Cell is far from being a homogeneous medium. This creates a number of problems which will be discussed as well as the ways to solve them. We will focus on the following problems: 1. Cellular surface brush (microvilli, glycocalyx..) surrounds cells. A simple model to separate the brush and cell deformation will be overviewed. 2. Problem of nonlinearity of stress-strain relation. Although unsolved, this may indirectly tested: the rigidity modulus should be reasonably independent of penetration. We will show that it can be achieved when using relatively dull AFM probes (the radii of microns) only. 3. Heterogeneity of cell surface. Cells are not homogeneous over the surface. We will discuss the question how many points is enough to characterize the cell. We will exemplify the above with human cervical epithelial cells. We will demonstrate that without proper consideration of the above problems, the error in defining the modulus of rigidity can easily reach an order of magnitude.

  2. Characterization of fiber-forming peptides and proteins by means of atomic force microscopy.

    PubMed

    Creasey, Rhiannon G; Gibson, Christopher T; Voelcker, Nicolas H

    2012-05-01

    The atomic force microscope (AFM) is widely used in biological sciences due to its ability to perform imaging experiments at high resolution in a physiological environment, without special sample preparation such as fixation or staining. AFM is unique, in that it allows single molecule information of mechanical properties and molecular recognition to be gathered. This review sets out to identify methodological applications of AFM for characterization of fiber-forming proteins and peptides. The basics of AFM operation are detailed, with in-depth information for any life scientist to get a grasp on AFM capabilities. It also briefly describes antibody recognition imaging and mapping of nanomechanical properties on biological samples. Subsequently, examples of AFM application to fiber-forming natural proteins, and fiber-forming synthetic peptides are given. Here, AFM is used primarily for structural characterization of fibers in combination with other techniques, such as circular dichroism and fluorescence spectroscopy. More recent developments in antibody recognition imaging to identify constituents of protein fibers formed in human disease are explored. This review, as a whole, seeks to encourage the life scientists dealing with protein aggregation phenomena to consider AFM as a part of their research toolkit, by highlighting the manifold capabilities of this technique.

  3. On the effects of intercalators in DNA condensation: a force spectroscopy and gel electrophoresis study.

    PubMed

    Rocha, M S; Cavalcante, A G; Silva, R; Ramos, E B

    2014-05-01

    In this work we have characterized the effects of the intercalator ethidium bromide (EtBr) on the DNA condensation process by using force spectroscopy and gel electrophoresis. We have tested two condensing agents: spermine (spm(4+)), a tetravalent cationic amine which promotes cation-induced DNA condensation, and poly(ethylene glycol) (PEG), a neutral polymer which promotes DNA ψ-condensation. Two different types of experiments were performed. In the first type, bare DNA molecules disperse in solution are first treated with EtBr for intercalation, and then the condensing agent is added to the sample with the purpose of verifying the effects of the intercalator in hindering DNA condensation. In the second experiment type, the bare DNA molecules are first condensed, and then the intercalator is added to the sample in order to verify its influence on the previously condensed DNA. The results obtained with the two different experimental techniques used agree very well, indicating that previously intercalated EtBr can hinder both cation-induced and ψ-condensation, being more efficient in the first case. On the other hand, EtBr has little effect on the previously formed cation-induced condensates, but is efficient in unfolding the ψ-condensates.

  4. Mechanisms of small molecule-DNA interactions probed by single-molecule force spectroscopy.

    PubMed

    Almaqwashi, Ali A; Paramanathan, Thayaparan; Rouzina, Ioulia; Williams, Mark C

    2016-05-19

    There is a wide range of applications for non-covalent DNA binding ligands, and optimization of such interactions requires detailed understanding of the binding mechanisms. One important class of these ligands is that of intercalators, which bind DNA by inserting aromatic moieties between adjacent DNA base pairs. Characterizing the dynamic and equilibrium aspects of DNA-intercalator complex assembly may allow optimization of DNA binding for specific functions. Single-molecule force spectroscopy studies have recently revealed new details about the molecular mechanisms governing DNA intercalation. These studies can provide the binding kinetics and affinity as well as determining the magnitude of the double helix structural deformations during the dynamic assembly of DNA-ligand complexes. These results may in turn guide the rational design of intercalators synthesized for DNA-targeted drugs, optical probes, or integrated biological self-assembly processes. Herein, we survey the progress in experimental methods as well as the corresponding analysis framework for understanding single molecule DNA binding mechanisms. We discuss briefly minor and major groove binding ligands, and then focus on intercalators, which have been probed extensively with these methods. Conventional mono-intercalators and bis-intercalators are discussed, followed by unconventional DNA intercalation. We then consider the prospects for using these methods in optimizing conventional and unconventional DNA-intercalating small molecules. PMID:27085806

  5. Mechanisms of small molecule–DNA interactions probed by single-molecule force spectroscopy

    PubMed Central

    Almaqwashi, Ali A.; Paramanathan, Thayaparan; Rouzina, Ioulia; Williams, Mark C.

    2016-01-01

    There is a wide range of applications for non-covalent DNA binding ligands, and optimization of such interactions requires detailed understanding of the binding mechanisms. One important class of these ligands is that of intercalators, which bind DNA by inserting aromatic moieties between adjacent DNA base pairs. Characterizing the dynamic and equilibrium aspects of DNA-intercalator complex assembly may allow optimization of DNA binding for specific functions. Single-molecule force spectroscopy studies have recently revealed new details about the molecular mechanisms governing DNA intercalation. These studies can provide the binding kinetics and affinity as well as determining the magnitude of the double helix structural deformations during the dynamic assembly of DNA–ligand complexes. These results may in turn guide the rational design of intercalators synthesized for DNA-targeted drugs, optical probes, or integrated biological self-assembly processes. Herein, we survey the progress in experimental methods as well as the corresponding analysis framework for understanding single molecule DNA binding mechanisms. We discuss briefly minor and major groove binding ligands, and then focus on intercalators, which have been probed extensively with these methods. Conventional mono-intercalators and bis-intercalators are discussed, followed by unconventional DNA intercalation. We then consider the prospects for using these methods in optimizing conventional and unconventional DNA-intercalating small molecules. PMID:27085806

  6. Investigating the mechanical properties of zona pellucida of whole human oocytes by atomic force spectroscopy.

    PubMed

    Andolfi, Laura; Masiero, Elena; Giolo, Elena; Martinelli, Monica; Luppi, Stefania; Dal Zilio, Simone; Delfino, Ines; Bortul, Roberta; Zweyer, Marina; Ricci, Giuseppe; Lazzarino, Marco

    2016-08-01

    The role of mechanics in numerous biological processes is nowadays recognized, while in others, such as the fertilization process, it is still neglected. In the case of oocytes the description of their mechanical properties could improve the comprehension of the oocyte-spermatozoon interaction and be helpful for application in in vitro fertilization (IVF) clinics. Herein the mechanical properties of whole human oocytes (HOs) immediately after retrieval are investigated by indentation measurements with atomic force spectroscopy under physiological conditions. Measurements are performed on immature (metaphase I - MI) and mature (metaphase II - MII) HOs. According to their morphological characteristics MII-HOs are classified as "suitable" and "rejected"; these latter would be usually rejected for intracytoplasmic sperm injection (ICSI). For all maturation stages we observe that the elastic response of the zona pellucida (ZP) outer layer was different and distinguishable from the rest of the ZP-HO. The elasticity of this ZP outer layer varies with maturation and quality: stiffness decreases from immature MI to good quality MII, up to poor-quality rejected MII. An indirect analysis with IVF outcome indicates that the ZP outer layer of analysed HOs donated by women who achieved pregnancy is stiffer than that of HOs from women with negative outcome. Our findings suggest that mechanical properties can represent important oocyte quality indicators that may be exploited for the design of innovative ICSI dedicated cell sorters. PMID:27476747

  7. Deciphering the energy landscape of the interaction uranyl-DCP with antibodies using dynamic force spectroscopy.

    PubMed

    Teulon, Jean-Marie; Parot, Pierre; Odorico, Michael; Pellequer, Jean-Luc

    2008-11-15

    Previous studies on molecular recognition of uranyl-DCP (dicarboxy-phenanthroline chelator) compound by two distinct monoclonal antibodies (Mabs U04S and U08S) clearly showed the presence of a biphasic shape in Bell-Evans' plots and an accentuated difference in slopes at the high loading rates. To further explore the basis in the slope difference, we have performed complementary experiments using antibody PHE03S, raised against uranyl-DCP but, presenting a strong cross-reactivity toward the DCP chelator. This work allowed us to obtain a reallocation of the respective contributions of the metal ion itself and that of the chelator. Results led us to propose a 2D schematic model representing two energy barriers observed in the systems Mabs U04S- and U08S-[UO(2)-DCP] where the outer barrier characterizes the interaction between UO(2) and Mab whereas the inner barrier characterizes the interaction between DCP and Mab. Using dynamic force spectroscopy, it is thus possible to dissect molecular interactions during the unbinding between proteins and ligands.

  8. MDM2-MDM4 molecular interaction investigated by atomic force spectroscopy and surface plasmon resonance.

    PubMed

    Moscetti, Ilaria; Teveroni, Emanuela; Moretti, Fabiola; Bizzarri, Anna Rita; Cannistraro, Salvatore

    2016-01-01

    Murine double minute 2 (MDM2) and 4 (MDM4) are known as the main negative regulators of p53, a tumor suppressor. They are able to form heterodimers that are much more effective in the downregulation of p53. Therefore, the MDM2-MDM4 complex could be a target for promising therapeutic restoration of p53 function. To this aim, a deeper understanding of the molecular mechanisms underlining the heterodimerization is needed. The kinetic and thermodynamic characterization of the MDM2-MDM4 complex was performed with two complementary approaches: atomic force spectroscopy and surface plasmon resonance. Both techniques revealed an equilibrium dissociation constant (KD ) in the micromolar range for the MDM2-MDM4 heterodimer, similar to related complexes involved in the p53 network. Furthermore, the MDM2-MDM4 complex is characterized by a relatively high free energy, through a single energy barrier, and by a lifetime in the order of tens of seconds. New insights into the MDM2-MDM4 interaction could be highly important for developing innovative anticancer drugs focused on p53 reactivation. PMID:27621617

  9. Biaxial Dielectrophoresis Force Spectroscopy: A Stoichiometric Approach for Examining Intermolecular Weak Binding Interactions.

    PubMed

    Park, In Soo; Kwak, Tae Joon; Lee, Gyudo; Son, Myeonggu; Choi, Jeong Woo; Choi, Seungyeop; Nam, Kihwan; Lee, Sei-Young; Chang, Woo-Jin; Eom, Kilho; Yoon, Dae Sung; Lee, Sangyoup; Bashir, Rashid; Lee, Sang Woo

    2016-04-26

    The direct quantification of weak intermolecular binding interactions is very important for many applications in biology and medicine. Techniques that can be used to investigate such interactions under a controlled environment, while varying different parameters such as loading rate, pulling direction, rupture event measurements, and the use of different functionalized probes, are still lacking. Herein, we demonstrate a biaxial dielectrophoresis force spectroscopy (BDFS) method that can be used to investigate weak unbinding events in a high-throughput manner under controlled environments and by varying the pulling direction (i.e., transverse and/or vertical axes) as well as the loading rate. With the BDFS system, we can quantitatively analyze binding interactions related to hydrogen bonding or ionic attractions between functionalized microbeads and a surface within a microfluidic device. Our BDFS system allowed for the characterization of the number of bonds involved in an interaction, bond affinity, kinetic rates, and energy barrier heights and widths from different regimes of the energy landscape. PMID:27007455

  10. Intrinsically high-Q dynamic AFM imaging in liquid with a significantly extended needle tip

    NASA Astrophysics Data System (ADS)

    Minary-Jolandan, Majid; Tajik, Arash; Wang, Ning; Yu, Min-Feng

    2012-06-01

    Atomic force microscope (AFM) probe with a long and rigid needle tip was fabricated and studied for high Q factor dynamic (tapping mode) AFM imaging of samples submersed in liquid. The extended needle tip over a regular commercially available tapping-mode AFM cantilever was sufficiently long to keep the AFM cantilever from submersed in liquid, which significantly minimized the hydrodynamic damping involved in dynamic AFM imaging of samples in liquid. Dynamic AFM imaging of samples in liquid at an intrinsic Q factor of over 100 and an operational frequency of over 200 kHz was demonstrated. The method has the potential to be extended to acquire viscoelastic material properties and provide truly gentle imaging of soft biological samples in physiological environments.

  11. Effect of tip mass on frequency response and sensitivity of AFM cantilever in liquid.

    PubMed

    Farokh Payam, Amir; Fathipour, Morteza

    2015-03-01

    The effect of tip mass on the frequency response and sensitivity of atomic force microscope (AFM) cantilever in the liquid environment is investigated. For this purpose, using Euler-Bernoulli beam theory and considering tip mass and hydrodynamic functions in a liquid environment, an expression for the resonance frequencies of AFM cantilever in liquid is derived. Then, based on this expression, the effect of the surface contact stiffness on the flexural mode of a rectangular AFM cantilever in fluid is investigated and compared with the case where the AFM cantilever operates in the air. The results show that in contrast with an air environment, the tip mass has no significant impact on the resonance frequency and sensitivity of the AFM cantilever in the liquid. Hence, analysis of AFM behaviour in liquid environment by neglecting the tip mass is logical. PMID:25562584

  12. AFM of biological complexes: what can we learn?

    PubMed Central

    Gaczynska, Maria; Osmulski, Pawel A.

    2009-01-01

    The term “biological complexes” broadly encompasses particles as diverse as multisubunit enzymes, viral capsids, transport cages, molecular nets, ribosomes, nucleosomes, biological membrane components and amyloids. The complexes represent a broad range of stability and composition. Atomic force microscopy offers a wealth of structural and functional data about such assemblies. For this review, we choose to comment on the significance of AFM to study various aspects of biology of selected nonmembrane protein assemblies. Such particles are large enough to reveal many structural details under the AFM probe. Importantly, the specific advantages of the method allow for gathering dynamic information about their formation, stability or allosteric structural changes critical for their function. Some of them have already found their way to nanomedical or nanotechnological applications. Here we present examples of studies where the AFM provided pioneering information about the biology of complexes, and examples of studies where the simplicity of the method is used toward the development of potential diagnostic applications. PMID:19802337

  13. Liquid contact resonance AFM: analytical models, experiments, and limitations

    NASA Astrophysics Data System (ADS)

    Parlak, Zehra; Tu, Qing; Zauscher, Stefan

    2014-11-01

    Contact resonance AFM (CR-AFM) is a scanning probe microscopy technique that utilizes the contact resonances of the AFM cantilever for concurrent imaging of topography and surface stiffness. The technique has not been used in liquid until recently due to analytical and experimental difficulties, associated with viscous damping of cantilever vibrations and fluid loading effects. To address these difficulties, (i) an analytical approach for contact resonances in liquid is developed, and (ii) direct excitation of the contact resonances is demonstrated by actuating the cantilever directly in a magnetic field. By implementing the analytical approach and the direct actuation through magnetic particles, quantitative stiffness imaging on surfaces with a wide range of stiffness can be achieved in liquid with soft cantilevers and low contact forces.

  14. Mounting of Escherichia coli spheroplasts for AFM imaging.

    SciTech Connect

    Sullivan, Claretta J; Morrell-Falvey, Jennifer L; Allison, David P; Doktycz, Mitchel John

    2005-11-01

    The cytoplasmic membrane of Escherichia coli (E. coli) is the location of numerous, chemically specific transporters and recognition elements. Investigation of this membrane in vivo by atomic force microscopy (AFM) requires removal of the cell wall and stable immobilization of the spheroplast. AFM images demonstrate that spheroplasts can be secured with warm gelatin applied to the mica substrate just before the addition of a spheroplast suspension. The resulting preparation can be repeatedly imaged by AFM over the course of several hours. Confocal fluorescence imaging confirms the association of the spheroplasts with the gelatin layer. Gelatin molecules are known to reorder into a network after heating. Entrapment within this gelatin network is believed to be responsible for the immobilization of spheroplasts on mica.

  15. Studying the mechanism of CD47-SIRPα interactions on red blood cells by single molecule force spectroscopy

    NASA Astrophysics Data System (ADS)

    Pan, Yangang; Wang, Feng; Liu, Yanhou; Jiang, Junguang; Yang, Yong-Guang; Wang, Hongda

    2014-08-01

    The interaction forces and binding kinetics between SIRPα and CD47 were investigated by single-molecule force spectroscopy (SMFS) on both fresh and experimentally aged human red blood cells (hRBCs). We found that CD47 experienced a conformation change after oxidation, which influenced the interaction force and the position of the energy barrier between SIRPα and CD47. Our results are significant for understanding the mechanism of phagocytosis of red blood cells at the single molecule level.The interaction forces and binding kinetics between SIRPα and CD47 were investigated by single-molecule force spectroscopy (SMFS) on both fresh and experimentally aged human red blood cells (hRBCs). We found that CD47 experienced a conformation change after oxidation, which influenced the interaction force and the position of the energy barrier between SIRPα and CD47. Our results are significant for understanding the mechanism of phagocytosis of red blood cells at the single molecule level. Electronic supplementary information (ESI) available: Experimental section. See DOI: 10.1039/c4nr02889a

  16. How does the molecular linker in dynamic force spectroscopy affect probing molecular interactions at the single-molecule level?

    NASA Astrophysics Data System (ADS)

    Taninaka, Atsushi; Aizawa, Kota; Hanyu, Tatsuya; Hirano, Yuuichi; Takeuchi, Osamu; Shigekawa, Hidemi

    2016-08-01

    Dynamic force spectroscopy (DFS) based on atomic force microscopy, which enables us to obtain information on the interaction potential between molecules such as antigen-antibody complexes at the single-molecule level, is a key technique for advancing molecular science and technology. However, to ensure the reliability of DFS measurement, its basic mechanism must be well understood. We examined the effect of the molecular linker used to fix the target molecule to the atomic force microscope cantilever, i.e., the force direction during measurement, for the first time, which has not been discussed until now despite its importance. The effect on the lifetime and barrier position, which can be obtained by DFS, was found to be ˜10 and ˜50%, respectively, confirming the high potential of DFS.

  17. How does the molecular linker in dynamic force spectroscopy affect probing molecular interactions at the single-molecule level?

    NASA Astrophysics Data System (ADS)

    Taninaka, Atsushi; Aizawa, Kota; Hanyu, Tatsuya; Hirano, Yuuichi; Takeuchi, Osamu; Shigekawa, Hidemi

    2016-08-01

    Dynamic force spectroscopy (DFS) based on atomic force microscopy, which enables us to obtain information on the interaction potential between molecules such as antigen–antibody complexes at the single-molecule level, is a key technique for advancing molecular science and technology. However, to ensure the reliability of DFS measurement, its basic mechanism must be well understood. We examined the effect of the molecular linker used to fix the target molecule to the atomic force microscope cantilever, i.e., the force direction during measurement, for the first time, which has not been discussed until now despite its importance. The effect on the lifetime and barrier position, which can be obtained by DFS, was found to be ∼10 and ∼50%, respectively, confirming the high potential of DFS.

  18. Structural investigations on native collagen type I fibrils using AFM

    SciTech Connect

    Strasser, Stefan; Zink, Albert; Janko, Marek; Heckl, Wolfgang M.; Thalhammer, Stefan . E-mail: stefan.thalhammer@gsf.de

    2007-03-02

    This study was carried out to determine the elastic properties of single collagen type I fibrils with the use of atomic force microscopy (AFM). Native collagen fibrils were formed by self-assembly in vitro characterized with the AFM. To confirm the inner assembly of the collagen fibrils, the AFM was used as a microdissection tool. Native collagen type I fibrils were dissected and the inner core uncovered. To determine the elastic properties of collagen fibrils the tip of the AFM was used as a nanoindentor by recording force-displacement curves. Measurements were done on the outer shell and in the core of the fibril. The structural investigations revealed the banding of the shell also in the core of native collagen fibrils. Nanoindentation experiments showed the same Young's modulus on the shell as well as in the core of the investigated native collagen fibrils. In addition, the measurements indicate a higher adhesion in the core of the collagen fibrils compared to the shell.

  19. Scanning hall probe microscopy (SHPM) using quartz crystal AFM feedback.

    PubMed

    Dede, M; Urkmen, K; Girişen, O; Atabak, M; Oral, A; Farrer, I; Ritchie, D

    2008-02-01

    Scanning Hall Probe Microscopy (SHPM) is a quantitative and non-invasive technique for imaging localized surface magnetic field fluctuations such as ferromagnetic domains with high spatial and magnetic field resolution of approximately 50 nm and 7 mG/Hz(1/2) at room temperature. In the SHPM technique, scanning tunneling microscope (STM) or atomic force microscope (AFM) feedback is used to keep the Hall sensor in close proximity of the sample surface. However, STM tracking SHPM requires conductive samples; therefore the insulating substrates have to be coated with a thin layer of gold. This constraint can be eliminated with the AFM feedback using sophisticated Hall probes that are integrated with AFM cantilevers. However it is very difficult to micro fabricate these sensors. In this work, we have eliminated the difficulty in the cantilever-Hall probe integration process, just by gluing a Hall Probe chip to a quartz crystal tuning fork force sensor. The Hall sensor chip is simply glued at the end of a 32.768 kHz or 100 kHz Quartz crystal, which is used as force sensor. An LT-SHPM system is used to scan the samples. The sensor assembly is dithered at the resonance frequency using a digital Phase Locked Loop circuit and frequency shifts are used for AFM tracking. SHPM electronics is modified to detect AFM topography and the frequency shift, along with the magnetic field image. Magnetic domains and topography of an Iron Garnet thin film crystal, NdFeB demagnetised magnet and hard disk samples are presented at room temperature. The performance is found to be comparable with the SHPM using STM feedback.

  20. Electrochemical oxidation of the chalcopyrite surface: an XPS and AFM study in solution at pH 4

    NASA Astrophysics Data System (ADS)

    Farquhar, Morag L.; Wincott, Paul L.; Wogelius, Roy A.; Vaughan, David J.

    2003-09-01

    The electrochemical oxidation of chalcopyrite (CuFeS 2) has been studied at pH 4 using voltammetry, coulometry, X-ray photoelectron spectroscopy (XPS) and both ex situ and in situ atomic force microscopy (AFM). Between 500 and 650 mV an anodic oxidation peak is observed, prior to the onset of the main decomposition reactions. Chalcopyrite electrodes in contact with electrolyte show some release of Cu into solution even without an applied potential. At 500 and 650 mV, the loss of Cu from the surface increases by a factor of 2 and 6, respectively. Oxidation at 500 mV results in the formation of a mixed oxide or hydroxide of iron, coincident with islands (<0.15 μm wide) of reaction products observed on the surface using AFM. The surface coverage of these islands increases with amount of charge passed. Oxidation at 650 mV shows similar processes have occurred, but with a greater island surface coverage and a more deeply altered surface. XPS depth profiling suggests iron oxide or hydroxide is now a major phase in the top ˜40 Å, with significant sulphate also formed. Observation of islands (alteration products) using in situ AFM under potential control shows that these features are not an artefact of the preparation methods.

  1. CD-AFM reference metrology at NIST and SEMATECH

    NASA Astrophysics Data System (ADS)

    Dixson, Ronald; Fu, Joseph; Orji, Ndubuisi; Guthrie, William; Allen, Richard; Cresswell, Michael

    2005-05-01

    The National Institute of Standards and Technology (NIST) and SEMATECH have been working together to improve the traceability of critical dimension atomic force microscope (CD-AFM) dimensional metrology in semiconductor manufacturing. A major component of this collaboration has been the implementation of a Reference Measurement System (RMS) at SEMATECH using a current generation CD-AFM. An earlier tool, originally used at SEMATECH, has now been installed at NIST. Uncertainty budgets were developed for pitch, height, and CD measurements using both tools. At present, the standard uncertainties are approximately 0.2 % for pitch measurements and 0.4% for step height measurements. Prior to the current work, CD AFM linewidth measurements were limited to a standard uncertainty of about 5 nm. However, this limit can now be significantly reduced. This reduction results from the completion of the NIST/SEMATECH collaboration on the development of single crystal critical dimension reference materials (SCDDRM). A new generation of these reference materials was released to SEMATECH Member Companies during late 2004. The SEMATECH RMS was used to measure the linewidths of selected features on the distributed specimens. To reduce the uncertainty in tip width calibration, a separate transfer experiment was performed in which samples were measured by CD-AFM and then sent for high resolution transmission electron microscopy (HRTEM). In this manner, CD-AFM could be used to transfer the HRTEM width information to the distributed samples. Consequently, we are now able to reduce the limit on the standard uncertainty (k = 1) of CD-AFM width measurements to 1 nm.

  2. Polymer coatings on conductive polypyrroles surface characterization by XPS, ToFSIMS, inverse gas chromatography and AFM

    SciTech Connect

    Chehimi, M.M.; Abel, M.; Delamar, M.; Watts, J.F.; Zhdan, P.A.

    1996-01-01

    The study of PMMA adsorption on some conducting polypyrroles (PPys) using a variety of surface analytical techniques is reported. PMMA adsorption was monitored by X-ray photoelectron spectroscopy (XPS), time of flight secondary ion mass spectroscopy (ToF-SIMS) and inverse gas chromatography (IGC). XPS and ToF-SIMS permit to determine the surface composition of PMMA-coated PPy surfaces vs the solvent nature, temperature and the PPy dopant anion. Both techniques show that acid-base interactions may govern PMMA adsorption. IGC was used to determine the coating morphology by monitoring the surface energy of the coated PPy powders. It is suggested that homogeneity of PMMA coatings increases with decreasing solvent power. Preliminary atomic force microscopy (AFM) results on PMMA films cast on flat PPy surfaces confirm the IGC observation. {copyright} {ital 1996 American Institute of Physics.}

  3. Model for Stretching and Unfolding the Giant Multidomain Muscle Protein Using Single-Molecule Force Spectroscopy

    NASA Astrophysics Data System (ADS)

    Staple, Douglas B.; Payne, Stephen H.; Reddin, Andrew L. C.; Kreuzer, Hans Jürgen

    2008-12-01

    Single-molecule manipulation has allowed the forced unfolding of multidomain proteins. Here we outline a theory that not only explains these experiments but also points out a number of difficulties in their interpretation and makes suggestions for further experiments. For titin we reproduce force-extension curves, the dependence of break force on pulling speed, and break-force distributions and also validate two common experimental views: Unfolding titin Ig domains can be explained as stepwise increases in contour length, and increasing force peaks in native Ig sequences represent a hierarchy of bond strengths. Our theory is valid for essentially any molecule that can be unfolded in atomic force microscopy; as a further example, we present force-extension curves for the unfolding of RNA hairpins.

  4. Modeling the effect of cell-associated polymeric fluid layers on force spectroscopy measurements. Part I: model development.

    PubMed

    Coldren, Faith M; Foteinopoulou, Katerina; Carroll, David L; Laso, Manuel

    2008-09-01

    The mechanical response, the force-indentation relationship, in normal force spectroscopy measurements carried out on individual polysaccharide encapsulated bacteria is modeled using three increasingly refined approaches that consider the elastic response of the bacterium and cantilever in combination with a fluid (hydrodynamic) model for the polysaccharide layer. For the hydrodynamic description of the polysaccharide layer, several increasingly realistic models are described in detail, together with numerical solution techniques. These models range from one-dimensional, Newtonian, to two-dimensional, axisymmetric, fully viscoelastic (Phan-Thien/Tanner). In all cases, the models rigorously consider the time-dependent rheological-mechanical coupling between the elastic and fluid viscoelastic physical components of the experimental setup. Effects of inherent variability in geometrical and material properties of the bacterium and polysaccharide layer on the measurable response are quantified. A parametric investigation of the force-indentation relationship highlights the importance of accurate knowledge of the rheology of the extracellular polysaccharides. We also draw conclusions about the design and evaluation of force spectroscopy experiments on single encapsulated bacteria. Supported by model calculations, we also point the way to methods of in vivo rheological characterization of the extracellular polysaccharide as a preferable alternative to characterization after its removal from the native environment. PMID:18666790

  5. Spectrin-ankyrin interaction mechanics: A key force balance factor in the red blood cell membrane skeleton.

    PubMed

    Saito, Masakazu; Watanabe-Nakayama, Takahiro; Machida, Shinichi; Osada, Toshiya; Afrin, Rehana; Ikai, Atsushi

    2015-01-01

    As major components of red blood cell (RBC) cytoskeleton, spectrin and F-actin form a network that covers the entire cytoplasmic surface of the plasma membrane. The cross-linked two layered structure, called the membrane skeleton, keeps the structural integrity of RBC under drastically changing mechanical environment during circulation. We performed force spectroscopy experiments on the atomic force microscope (AFM) as a means to clarify the mechanical characteristics of spectrin-ankyrin interaction, a key factor in the force balance of the RBC cytoskeletal structure. An AFM tip was functionalized with ANK1-62k and used to probe spectrin crosslinked to mica surface. A force spectroscopy study gave a mean unbinding force of ~30 pN under our experimental conditions. Two energy barriers were identified in the unbinding process. The result was related to the well-known flexibility of spectrin tetramer and participation of ankyrin 1-spectrin interaction in the overall balance of membrane skeleton dynamics.

  6. Single molecule force spectroscopy data and BD- and MD simulations on the blood protein von Willebrand factor.

    PubMed

    Posch, Sandra; Aponte-Santamaría, Camilo; Schwarzl, Richard; Karner, Andreas; Radtke, Matthias; Gräter, Frauke; Obser, Tobias; König, Gesa; Brehm, Maria A; Gruber, Hermann J; Netz, Roland R; Baldauf, Carsten; Schneppenheim, Reinhard; Tampé, Robert; Hinterdorfer, Peter

    2016-09-01

    We here give information for a deeper understanding of single molecule force spectroscopy (SMFS) data through the example of the blood protein von Willebrand factor (VWF). It is also shown, how fitting of rupture forces versus loading rate profiles in the molecular dynamics (MD) loading-rate range can be used to demonstrate the qualitative agreement between SMFS and MD simulations. The recently developed model by Bullerjahn, Sturm, and Kroy (BSK) was used for this demonstration. Further, Brownian dynamics (BD) simulations, which can be utilized to estimate the lifetimes of intramolecular VWF interactions under physiological shear, are described. For interpretation and discussion of the methods and data presented here, we would like to directly point the reader to the related research paper, "Mutual A domain interactions in the force sensing protein von Willebrand Factor" (Posch et al., 2016) [1]. PMID:27508268

  7. Single molecule force spectroscopy data and BD- and MD simulations on the blood protein von Willebrand factor.

    PubMed

    Posch, Sandra; Aponte-Santamaría, Camilo; Schwarzl, Richard; Karner, Andreas; Radtke, Matthias; Gräter, Frauke; Obser, Tobias; König, Gesa; Brehm, Maria A; Gruber, Hermann J; Netz, Roland R; Baldauf, Carsten; Schneppenheim, Reinhard; Tampé, Robert; Hinterdorfer, Peter

    2016-09-01

    We here give information for a deeper understanding of single molecule force spectroscopy (SMFS) data through the example of the blood protein von Willebrand factor (VWF). It is also shown, how fitting of rupture forces versus loading rate profiles in the molecular dynamics (MD) loading-rate range can be used to demonstrate the qualitative agreement between SMFS and MD simulations. The recently developed model by Bullerjahn, Sturm, and Kroy (BSK) was used for this demonstration. Further, Brownian dynamics (BD) simulations, which can be utilized to estimate the lifetimes of intramolecular VWF interactions under physiological shear, are described. For interpretation and discussion of the methods and data presented here, we would like to directly point the reader to the related research paper, "Mutual A domain interactions in the force sensing protein von Willebrand Factor" (Posch et al., 2016) [1].

  8. Three-dimensional interaction force and tunneling current spectroscopy of point defects on rutile TiO2(110)

    NASA Astrophysics Data System (ADS)

    Baykara, Mehmet Z.; Mönig, Harry; Schwendemann, Todd C.; Ünverdi, Ã.-zhan; Altman, Eric I.; Schwarz, Udo D.

    2016-02-01

    The extent to which point defects affect the local chemical reactivity and electronic properties of an oxide surface was evaluated with picometer resolution in all three spatial dimensions using simultaneous atomic force/scanning tunneling microscopy measurements performed on the (110) face of rutile TiO2. Oxygen atoms were imaged as protrusions in both data channels, corresponding to a rarely observed imaging mode for this prototypical metal oxide surface. Three-dimensional spectroscopy of interaction forces and tunneling currents was performed on individual surface and subsurface defects as a function of tip-sample distance. An interstitial defect assigned to a subsurface hydrogen atom is found to have a distinct effect on the local density of electronic states on the surface, but no detectable influence on the tip-sample interaction force. Meanwhile, spectroscopic data acquired on an oxygen vacancy highlight the role of the probe tip in chemical reactivity measurements.

  9. Atomic force microscopy and x-ray photoelectron spectroscopy investigations of the morphology and chemistry of a PdCl{sub 2}/SnCl{sub 2} electroless plating catalysis system adsorbed onto shape memory alloy particles

    SciTech Connect

    Silvain, J.F.; Fouassier, O.; Lescaux, S.

    2004-11-01

    A study of the different stages of the electroless deposition of copper on micronic NiTi shape memory alloy particles activated by one-step and two-step methods has been conducted from both a chemical and a morphological point of view. The combination of x-ray photoelectron spectroscopy (XPS) measurements and atomic force microscopy (AFM) imaging has allowed detection of the distribution of the formed compounds and depth quantification and estimation of the surface topographic parameters. For the two-step method, at the sensitization of the early stages, it is observed by AFM that Sn is absorbed in form of clusters that tend to completely cover the surface and form a continuous film. XPS analysis have shown that Sn and Pd are first absorbed in form of oxide (SnO{sub 2} and PdO) and hydroxide [Sn(OH){sub 4}]. After the entire sensitization step, the NiTi substrate is covered with Sn-based compounds. After the sensitization and the activation steps the powder roughness increases. Behavior of the Sn and Pd growth for the one-step method does not follow the behavior found for the two-step method. Indeed, XPS analysis shows a three-dimensional (3D) growth of Pd clusters on top of a mixture of metallic tin, oxide (SnO) and hydroxide [Sn(OH){sub 2}]. These Pd clusters are covered with a thin layer of Pd-oxide contamination induced by the electroless process. The mean roughness for the one-step and two-step processes are equivalent. After copper deposition, the decrease of mean roughness is attributed to a filling of surface valleys, observed after the Sn-Pd coating step.

  10. High-speed AFM for Studying Dynamic Biomolecular Processes

    NASA Astrophysics Data System (ADS)

    Ando, Toshio

    2008-03-01

    Biological molecules show their vital activities only in aqueous solutions. It had been one of dreams in biological sciences to directly observe biological macromolecules (protein, DNA) at work under a physiological condition because such observation is straightforward to understanding their dynamic behaviors and functional mechanisms. Optical microscopy has no sufficient spatial resolution and electron microscopy is not applicable to in-liquid samples. Atomic force microscopy (AFM) can visualize molecules in liquids at high resolution but its imaging rate was too low to capture dynamic biological processes. This slow imaging rate is because AFM employs mechanical probes (cantilevers) and mechanical scanners to detect the sample height at each pixel. It is quite difficult to quickly move a mechanical device of macroscopic size with sub-nanometer accuracy without producing unwanted vibrations. It is also difficult to maintain the delicate contact between a probe tip and fragile samples. Two key techniques are required to realize high-speed AFM for biological research; fast feedback control to maintain a weak tip-sample interaction force and a technique to suppress mechanical vibrations of the scanner. Various efforts have been carried out in the past decade to materialize high-speed AFM. The current high-speed AFM can capture images on video at 30-60 frames/s for a scan range of 250nm and 100 scan lines, without significantly disturbing week biomolecular interaction. Our recent studies demonstrated that this new microscope can reveal biomolecular processes such as myosin V walking along actin tracks and association/dissociation dynamics of chaperonin GroEL-GroES that occurs in a negatively cooperative manner. The capacity of nanometer-scale visualization of dynamic processes in liquids will innovate on biological research. In addition, it will open a new way to study dynamic chemical/physical processes of various phenomena that occur at the liquid-solid interfaces.

  11. Dissolution of uranophane: An AFM, XPS, SEM and ICP study

    NASA Astrophysics Data System (ADS)

    Schindler, Michael; Freund, Michael; Hawthorne, Frank C.; Burns, Peter C.; Maurice, Patricia A.

    2009-05-01

    Dissolution experiments on single crystals of uranophane and uranophane-β, Ca(H 2O) 5[(UO 2)(SiO 3(OH)] 2, from the Shinkolobwe mine of the Democratic Republic of Congo, were done in an aqueous HCl solution of pH 3.5 for 3 h, in HCl solutions of pH 2 for 5, 10 and 30 min, and in Pb 2+-, Ba-, Sr-, Ca- and Mg-HCl solutions of pH 2 for 30 min. The basal surfaces of the treated uranophane crystals were examined using atomic-force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM). Solutions after dissolution experiments on single crystals and synthetic powders were analysed with inductively coupled plasma-optical emission spectroscopy (ICP-OES) and mass spectroscopy (ICP-MS). The morphology of the observed etch pits (measured by AFM) were compared to the morphology, predicted on the basis of the bond-valence deficiency of polyhedron chains along the edges of the basal surface. Etch pits form in HCl solutions of pH 2. Their decrease in depth with the duration of the dissolution experiment is explained with the stepwave dissolution model, which describes the lowering of the surrounding area of an etch pit with continuous waves of steps emanated from the etch pit into the rest of the crystal surface. Hillocks form in an HCl solution of pH 3.5, and the chemical composition of the surface (as indicated by XPS) shows that these hillocks are the result of the precipitation of a uranyl-hydroxy-hydrate phase. Well-orientated hillocks form on the surface of uranophane in a SrCl 2-HCl solution of pH 2. They are part of an aged silica coating of composition Si 2O 2(OH) 4(H 2O) n. An amorphous layer forms on the surface of uranophane in a MgCl 2-HCl solution of pH 2, which has a composition and structure similar to silicic acid. Small crystallites of uranyl-hydroxy-hydrate phases form on the surface of uranophane after treatment in Pb(NO 3) 2-HCl and BaCl 2-HCl solutions of pH 2. Dissolution experiments on synthetic uranophane powders

  12. β-connectin studies by small-angle x-ray scattering and single-molecule force spectroscopy by atomic force microscopy

    NASA Astrophysics Data System (ADS)

    Marchetti, S.; Sbrana, F.; Toscano, A.; Fratini, E.; Carlà, M.; Vassalli, M.; Tiribilli, B.; Pacini, A.; Gambi, C. M. C.

    2011-05-01

    The three-dimensional structure and the mechanical properties of a β-connectin fragment from human cardiac muscle, belonging to the I band, from I27 to I34, were investigated by small-angle x-ray scattering (SAXS) and single-molecule force spectroscopy (SMFS). This molecule presents an entropic elasticity behavior, associated to globular domain unfolding, that has been widely studied in the last 10 years. In addition, atomic force microscopy based SMFS experiments suggest that this molecule has an additional elastic regime, for low forces, probably associated to tertiary structure remodeling. From a structural point of view, this behavior is a mark of the fact that the eight domains in the I27-I34 fragment are not independent and they organize in solution, assuming a well-defined three-dimensional structure. This hypothesis has been confirmed by SAXS scattering, both on a diluted and a concentrated sample. Two different models were used to fit the SAXS curves: one assuming a globular shape and one corresponding to an elongated conformation, both coupled with a Coulomb repulsion potential to take into account the protein-protein interaction. Due to the predominance of the structure factor, the effective shape of the protein in solution could not be clearly disclosed. By performing SMFS by atomic force microscopy, mechanical unfolding properties were investigated. Typical sawtooth profiles were obtained and the rupture force of each unfolding domain was estimated. By fitting a wormlike chain model to each peak of the sawtooth profile, the entropic elasticity of octamer was described.

  13. {beta}-connectin studies by small-angle x-ray scattering and single-molecule force spectroscopy by atomic force microscopy

    SciTech Connect

    Marchetti, S.; Carla, M.; Gambi, C. M. C.; Sbrana, F.; Vassalli, M.; Toscano, A.; Pacini, A.; Fratini, E.; Tiribilli, B.

    2011-05-15

    The three-dimensional structure and the mechanical properties of a {beta}-connectin fragment from human cardiac muscle, belonging to the I band, from I{sub 27} to I{sub 34}, were investigated by small-angle x-ray scattering (SAXS) and single-molecule force spectroscopy (SMFS). This molecule presents an entropic elasticity behavior, associated to globular domain unfolding, that has been widely studied in the last 10 years. In addition, atomic force microscopy based SMFS experiments suggest that this molecule has an additional elastic regime, for low forces, probably associated to tertiary structure remodeling. From a structural point of view, this behavior is a mark of the fact that the eight domains in the I{sub 27}-I{sub 34} fragment are not independent and they organize in solution, assuming a well-defined three-dimensional structure. This hypothesis has been confirmed by SAXS scattering, both on a diluted and a concentrated sample. Two different models were used to fit the SAXS curves: one assuming a globular shape and one corresponding to an elongated conformation, both coupled with a Coulomb repulsion potential to take into account the protein-protein interaction. Due to the predominance of the structure factor, the effective shape of the protein in solution could not be clearly disclosed. By performing SMFS by atomic force microscopy, mechanical unfolding properties were investigated. Typical sawtooth profiles were obtained and the rupture force of each unfolding domain was estimated. By fitting a wormlike chain model to each peak of the sawtooth profile, the entropic elasticity of octamer was described.

  14. XPS and AFM Study of GaAs Surface Treatment

    SciTech Connect

    Contreras-Guerrero, R.; Wallace, R. M.; Aguirre-Francisco, S.; Herrera-Gomez, A.; Lopez-Lopez, M.

    2008-11-13

    Obtaining smooth and atomically clean surfaces is an important step in the preparation of a surface for device manufacturing. In this work different processes are evaluated for cleaning a GaAs surface. A good surface cleaning treatment is that which provides a high level of uniformity and controllability of the surface. Different techniques are useful as cleaning treatments depending on the growth process to be used. The goal is to remove the oxygen and carbon contaminants and then form a thin oxide film to protect the surface, which is easy to remove later with thermal desorption mechanism like molecular beam epitaxy (MBE) with minimal impact to the surface. In this study, atomic force microscopy (AFM), x-ray photoelectron spectroscopy (XPS) and secondary ion mass spectrometry (SIMS) were used to characterize the structure of the surface, the composition, as well as detect oxygen and carbon contaminant on the GaAs surface. This study consists in two parts. The first part the surface was subjected to different chemical treatments. The chemical solutions were: (a)H{sub 2}SO{sub 4}:H{sub 2}O{sub 2}:H{sub 2}O(4:1:100), (b) HCl: H{sub 2}O(1:3), (c)NH{sub 4}OH 29%. The treatments (a) and (b) reduced the oxygen on the surface. Treatment (c) reduces carbon contamination. In the second part we made MOS devices on the surfaces treated. They were characterized by CV and IV electrical measurements. They show frequency dispersion.

  15. Using 2D Correlation Analysis to Enhance Spectral Information Available from Highly Spatially Resolved AFM-IR Spectra.

    PubMed

    Marcott, Curtis; Lo, Michael; Hu, Qichi; Kjoller, Kevin; Boskey, Adele; Noda, Isao

    2014-07-01

    The recent combination of atomic force microscopy and infrared spectroscopy (AFM-IR) has led to the ability to obtain IR spectra with nanoscale spatial resolution, nearly two orders-of-magnitude better than conventional Fourier transform infrared (FT-IR) microspectroscopy. This advanced methodology can lead to significantly sharper spectral features than are typically seen in conventional IR spectra of inhomogeneous materials, where a wider range of molecular environments are coaveraged by the larger sample cross section being probed. In this work, two-dimensional (2D) correlation analysis is used to examine position sensitive spectral variations in datasets of closely spaced AFM-IR spectra. This analysis can reveal new key insights, providing a better understanding of the new spectral information that was previously hidden under broader overlapped spectral features. Two examples of the utility of this new approach are presented. Two-dimensional correlation analysis of a set of AFM-IR spectra were collected at 200-nm increments along a line through a nucleation site generated by remelting a small spot on a thin film of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate). There are two different crystalline carbonyl band components near 1720 cm(-1) that sequentially disappear before a band at 1740 cm(-1) due to more disordered material appears. In the second example, 2D correlation analysis of a series of AFM-IR spectra spaced every 1 micrometer of a thin cross section of a bone sample measured outward from an osteon center of bone growth. There are many changes in the amide I and phosphate band contours, suggesting changes in the bone structure are occurring as the bone matures.

  16. Resonance Frequency Analysis for Surface-Coupled AFM Cantilever in Liquids

    SciTech Connect

    Mirman, B; Kalinin, Sergei V

    2008-01-01

    Shifts in the resonance frequencies of surface-coupled atomic force microscope (AFM) probes are used as the basis for the detection mechanisms in a number of scanning probe microscopy techniques including atomic force acoustic microscopy (AFAM), force modulation microscopy, and resonance enhanced piezoresponse force microscopy (PFM). Here, we analyze resonance characteristics for AFM cantilever coupled to surface in liquid environment, and derive approximate expressions for resonant frequencies as a function of vertical and lateral spring constant of the tip-surface junction. This analysis provides a simplified framework for the interpretation of AFAM and PFM data in ambient, liquid, and vacuum environments.

  17. Forces and Dynamics of Glucose and Inhibitor Binding to Sodium Glucose Co-transporter SGLT1 Studied by Single Molecule Force Spectroscopy*

    PubMed Central

    Neundlinger, Isabel; Puntheeranurak, Theeraporn; Wildling, Linda; Rankl, Christian; Wang, Lai-Xi; Gruber, Hermann J.; Kinne, Rolf K. H.; Hinterdorfer, Peter

    2014-01-01

    Single molecule force spectroscopy was employed to investigate the dynamics of the sodium glucose co-transporter (SGLT1) upon substrate and inhibitor binding on the single molecule level. CHO cells stably expressing rbSGLT1 were probed by using atomic force microscopy tips carrying either thioglucose, 2′-aminoethyl β-d-glucopyranoside, or aminophlorizin. Poly(ethylene glycol) (PEG) chains of different length and varying end groups were used as tether. Experiments were performed at 10, 25 and 37 °C to address different conformational states of SGLT1. Unbinding forces between ligands and SGLT1 were recorded at different loading rates by changing the retraction velocity, yielding binding probability, width of energy barrier of the binding pocket, and the kinetic off rate constant of the binding reaction. With increasing temperature, width of energy barrier and average life time increased for the interaction of SGLT1 with thioglucose (coupled via acrylamide to a long PEG) but decreased for aminophlorizin binding. The former indicates that in the membrane-bound SGLT1 the pathway to sugar translocation involves several steps with different temperature sensitivity. The latter suggests that also the aglucon binding sites for transport inhibitors have specific, temperature-sensitive conformations. PMID:24962566

  18. Evolution of nano-rheological properties of Nafion¯ thin films during pH modification by strong base treatment: A static and dynamic force spectroscopy study

    NASA Astrophysics Data System (ADS)

    Eslami, Babak; López-Guerra, Enrique A.; Raftari, Maryam; Solares, Santiago D.

    2016-04-01

    Addition of a strong base to Nafion® proton exchange membranes is a common practice in industry to increase their overall performance in fuel cells. Here, we investigate the evolution of the nano-rheological properties of Nafion thin films as a function of the casting pH, via characterization with static and dynamic, contact and intermittent-contact atomic force microscopy (AFM) techniques. The addition of KOH causes non-monotonic changes in the viscoelastic properties of the films, which behave as highly dissipative, softer materials near neutral pH values, and as harder, more elastic materials at extreme pH values. We quantify this behavior through calculation of the temporal evolution of the compliance and the glassy compliance under static AFM measurements. We complement these observations with dynamic AFM metrics, including dissipated power and virial (for intermittent-contact-mode measurements), and contact resonance frequency and quality factor (for dynamic contact-mode measurements). We explain the non-monotonic material property behavior in terms of the degree of ionic crosslinking and moisture content of the films, which vary with the addition of KOH. This work focuses on the special case study of the addition of strong bases, but the observed mechanical property changes are broadly related to water plasticizing effects and ionic crosslinking, which are also important in other types of films.

  19. 3D Color Digital Elevation Map of AFM Sample

    NASA Technical Reports Server (NTRS)

    2008-01-01

    This color image is a three dimensional (3D) view of a digital elevation map of a sample collected by NASA's Phoenix Mars Lander's Atomic Force Microscope (AFM).

    The image shows four round pits, only 5 microns in depth, that were micromachined into the silicon substrate, which is the background plane shown in red. This image has been processed to reflect the levelness of the substrate.

    A Martian particle only one micrometer, or one millionth of a meter, across is held in the upper left pit.

    The rounded particle shown at the highest magnification ever seen from another world is a particle of the dust that cloaks Mars. Such dust particles color the Martian sky pink, feed storms that regularly envelop the planet and produce Mars' distinctive red soil.

    The particle was part of a sample informally called 'Sorceress' delivered to the AFM on the 38th Martian day, or sol, of the mission (July 2, 2008). The AFM is part of Phoenix's microscopic station called MECA, or the Microscopy, Electrochemistry, and Conductivity Analyzer.

    The AFM was developed by a Swiss-led consortium, with Imperial College London producing the silicon substrate that holds sampled particles.

    The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  20. Single-molecule Force Spectroscopy Predicts a Misfolded, Domain-swapped Conformation in human γD-Crystallin Protein*

    PubMed Central

    Garcia-Manyes, Sergi; Giganti, David; Badilla, Carmen L.; Lezamiz, Ainhoa; Perales-Calvo, Judit; Beedle, Amy E. M.; Fernández, Julio M.

    2016-01-01

    Cataract is a protein misfolding disease where the size of the aggregate is directly related to the severity of the disorder. However, the molecular mechanisms that trigger the onset of aggregation remain unknown. Here we use a combination of protein engineering techniques and single-molecule force spectroscopy using atomic force microscopy to study the individual unfolding pathways of the human γD-crystallin, a multidomain protein that must remain correctly folded during the entire lifetime to guarantee lens transparency. When stretching individual polyproteins containing two neighboring HγD-crystallin monomers, we captured an anomalous misfolded conformation in which the β1 and β2 strands of the N terminus domain of two adjacent monomers swap. This experimentally elusive domain-swapped conformation is likely to be responsible for the increase in molecular aggregation that we measure in vitro. Our results demonstrate the power of force spectroscopy at capturing rare misfolded conformations with potential implications for the understanding of the molecular onset of protein aggregation. PMID:26703476

  1. A Multifunctional Frontloading Approach for Repeated Recycling of a Pressure-Controlled AFM Micropipette

    PubMed Central

    Roder, Phillip; Hille, Carsten

    2015-01-01

    Fluid force microscopy combines the positional accuracy and force sensitivity of an atomic force microscope (AFM) with nanofluidics via a microchanneled cantilever. However, adequate loading and cleaning procedures for such AFM micropipettes are required for various application situations. Here, a new frontloading procedure is described for an AFM micropipette functioning as a force- and pressure-controlled microscale liquid dispenser. This frontloading procedure seems especially attractive when using target substances featuring high costs or low available amounts. Here, the AFM micropipette could be filled from the tip side with liquid from a previously applied droplet with a volume of only a few μL using a short low-pressure pulse. The liquid-loaded AFM micropipettes could be then applied for experiments in air or liquid environments. AFM micropipette frontloading was evaluated with the well-known organic fluorescent dye rhodamine 6G and the AlexaFluor647-labeled antibody goat anti-rat IgG as an example of a larger biological compound. After micropipette usage, specific cleaning procedures were tested. Furthermore, a storage method is described, at which the AFM micropipettes could be stored for a few hours up to several days without drying out or clogging of the microchannel. In summary, the rapid, versatile and cost-efficient frontloading and cleaning procedure for the repeated usage of a single AFM micropipette is beneficial for various application situations from specific surface modifications through to local manipulation of living cells, and provides a simplified and faster handling for already known experiments with fluid force microscopy. PMID:26636981

  2. A Multifunctional Frontloading Approach for Repeated Recycling of a Pressure-Controlled AFM Micropipette.

    PubMed

    Roder, Phillip; Hille, Carsten

    2015-01-01

    Fluid force microscopy combines the positional accuracy and force sensitivity of an atomic force microscope (AFM) with nanofluidics via a microchanneled cantilever. However, adequate loading and cleaning procedures for such AFM micropipettes are required for various application situations. Here, a new frontloading procedure is described for an AFM micropipette functioning as a force- and pressure-controlled microscale liquid dispenser. This frontloading procedure seems especially attractive when using target substances featuring high costs or low available amounts. Here, the AFM micropipette could be filled from the tip side with liquid from a previously applied droplet with a volume of only a few μL using a short low-pressure pulse. The liquid-loaded AFM micropipettes could be then applied for experiments in air or liquid environments. AFM micropipette frontloading was evaluated with the well-known organic fluorescent dye rhodamine 6G and the AlexaFluor647-labeled antibody goat anti-rat IgG as an example of a larger biological compound. After micropipette usage, specific cleaning procedures were tested. Furthermore, a storage method is described, at which the AFM micropipettes could be stored for a few hours up to several days without drying out or clogging of the microchannel. In summary, the rapid, versatile and cost-efficient frontloading and cleaning procedure for the repeated usage of a single AFM micropipette is beneficial for various application situations from specific surface modifications through to local manipulation of living cells, and provides a simplified and faster handling for already known experiments with fluid force microscopy. PMID:26636981

  3. On CD-AFM bias related to probe bending

    NASA Astrophysics Data System (ADS)

    Ukraintsev, V. A.; Orji, N. G.; Vorburger, T. V.; Dixson, R. G.; Fu, J.; Silver, R. M.

    2012-03-01

    Critical Dimension AFM (CD-AFM) is a widely used reference metrology. To characterize modern semiconductor devices, very small and flexible probes, often 15 nm to 20 nm in diameter, are now frequently used. Several recent publications have reported on uncontrolled and significant probe-to-probe bias variation during linewidth and sidewall angle measurements [1,2]. Results obtained in this work suggest that probe bending can be on the order of several nanometers and thus potentially can explain much of the observed CD-AFM probe-to-probe bias variation. We have developed and experimentally tested one-dimensional (1D) and two-dimensional (2D) models to describe the bending of cylindrical probes. An earlier 1D bending model reported by Watanabe et al. [3] was refined. Contributions from several new phenomena were considered, including: probe misalignment, diameter variation near the carbon nanotube tip (CNT) apex, probe bending before snapping, distributed van der Waals-London force, etc. The methodology for extraction of the Hamaker probe-surface interaction energy from experimental probe bending data was developed. To overcome limitations of the 1D model, a new 2D distributed force (DF) model was developed. Comparison of the new model with the 1D single point force (SPF) model revealed about 27 % difference in probe bending bias between the two. A simple linear relation between biases predicted by the 1D SPF and 2D DF models was found. This finding simplifies use of the advanced 2D DF model of probe bending in various CD-AFM applications. New 2D and three-dimensional (3D) CDAFM data analysis software is needed to take full advantage of the new bias correction modeling capabilities.

  4. Topographical and Chemical Imaging of a Phase Separated Polymer Using a Combined Atomic Force Microscopy/Infrared Spectroscopy/Mass Spectrometry Platform

    DOE PAGES

    Tai, Tamin; Karácsony, Orsolya; Bocharova, Vera; Van Berkel, Gary J.; Kertesz, Vilmos

    2016-02-18

    This article describes how the use of a hybrid atomic force microscopy/infrared spectroscopy/mass spectrometry imaging platform was demonstrated for the acquisition and correlation of nanoscale sample surface topography and chemical images based on infrared spectroscopy and mass spectrometry.

  5. Theoretical modelling of AFM for bimetallic tip-substrate interactions

    NASA Technical Reports Server (NTRS)

    Bozzolo, Guillermo; Ferrante, John

    1991-01-01

    Recently, a new technique for calculating the defect energetics of alloys based on Equivalent Crystal Theory was developed. This new technique successfully predicts the bulk properties for binary alloys as well as segregation energies in the dilute limit. The authors apply this limit for the calculation of energy and force as a function of separation of an atomic force microscope (AFM) tip and substrate. The study was done for different combinations of tip and sample materials. The validity of the universality discovered for the same metal interfaces is examined for the case of different metal interactions.

  6. Life in extreme environments: single molecule force spectroscopy as a tool to explore proteins from extremophilic organisms.

    PubMed

    Tych, Katarzyna M; Hoffmann, Toni; Batchelor, Matthew; Hughes, Megan L; Kendrick, Katherine E; Walsh, Danielle L; Wilson, Michael; Brockwell, David J; Dougan, Lorna

    2015-04-01

    Extremophiles are organisms which survive and thrive in extreme environments. The proteins from extremophilic single-celled organisms have received considerable attention as they are structurally stable and functionally active under extreme physical and chemical conditions. In this short article, we provide an introduction to extremophiles, the structural adaptations of proteins from extremophilic organisms and the exploitation of these proteins in industrial applications. We provide a review of recent developments which have utilized single molecule force spectroscopy to mechanically manipulate proteins from extremophilic organisms and the information which has been gained about their stability, flexibility and underlying energy landscapes.

  7. A simple and efficient quasi 3-dimensional viscoelastic model and software for simulation of tapping-mode atomic force microscopy

    DOE PAGES

    Solares, Santiago D.

    2015-11-26

    This study introduces a quasi-3-dimensional (Q3D) viscoelastic model and software tool for use in atomic force microscopy (AFM) simulations. The model is based on a 2-dimensional array of standard linear solid (SLS) model elements. The well-known 1-dimensional SLS model is a textbook example in viscoelastic theory but is relatively new in AFM simulation. It is the simplest model that offers a qualitatively correct description of the most fundamental viscoelastic behaviors, namely stress relaxation and creep. However, this simple model does not reflect the correct curvature in the repulsive portion of the force curve, so its application in the quantitative interpretationmore » of AFM experiments is relatively limited. In the proposed Q3D model the use of an array of SLS elements leads to force curves that have the typical upward curvature in the repulsive region, while still offering a very low computational cost. Furthermore, the use of a multidimensional model allows for the study of AFM tips having non-ideal geometries, which can be extremely useful in practice. Examples of typical force curves are provided for single- and multifrequency tappingmode imaging, for both of which the force curves exhibit the expected features. Lastly, a software tool to simulate amplitude and phase spectroscopy curves is provided, which can be easily modified to implement other controls schemes in order to aid in the interpretation of AFM experiments.« less

  8. A simple and efficient quasi 3-dimensional viscoelastic model and software for simulation of tapping-mode atomic force microscopy

    SciTech Connect

    Solares, Santiago D.

    2015-11-26

    This study introduces a quasi-3-dimensional (Q3D) viscoelastic model and software tool for use in atomic force microscopy (AFM) simulations. The model is based on a 2-dimensional array of standard linear solid (SLS) model elements. The well-known 1-dimensional SLS model is a textbook example in viscoelastic theory but is relatively new in AFM simulation. It is the simplest model that offers a qualitatively correct description of the most fundamental viscoelastic behaviors, namely stress relaxation and creep. However, this simple model does not reflect the correct curvature in the repulsive portion of the force curve, so its application in the quantitative interpretation of AFM experiments is relatively limited. In the proposed Q3D model the use of an array of SLS elements leads to force curves that have the typical upward curvature in the repulsive region, while still offering a very low computational cost. Furthermore, the use of a multidimensional model allows for the study of AFM tips having non-ideal geometries, which can be extremely useful in practice. Examples of typical force curves are provided for single- and multifrequency tappingmode imaging, for both of which the force curves exhibit the expected features. Lastly, a software tool to simulate amplitude and phase spectroscopy curves is provided, which can be easily modified to implement other controls schemes in order to aid in the interpretation of AFM experiments.

  9. A simple and efficient quasi 3-dimensional viscoelastic model and software for simulation of tapping-mode atomic force microscopy.

    PubMed

    Solares, Santiago D

    2015-01-01

    This paper introduces a quasi-3-dimensional (Q3D) viscoelastic model and software tool for use in atomic force microscopy (AFM) simulations. The model is based on a 2-dimensional array of standard linear solid (SLS) model elements. The well-known 1-dimensional SLS model is a textbook example in viscoelastic theory but is relatively new in AFM simulation. It is the simplest model that offers a qualitatively correct description of the most fundamental viscoelastic behaviors, namely stress relaxation and creep. However, this simple model does not reflect the correct curvature in the repulsive portion of the force curve, so its application in the quantitative interpretation of AFM experiments is relatively limited. In the proposed Q3D model the use of an array of SLS elements leads to force curves that have the typical upward curvature in the repulsive region, while still offering a very low computational cost. Furthermore, the use of a multidimensional model allows for the study of AFM tips having non-ideal geometries, which can be extremely useful in practice. Examples of typical force curves are provided for single- and multifrequency tapping-mode imaging, for both of which the force curves exhibit the expected features. Finally, a software tool to simulate amplitude and phase spectroscopy curves is provided, which can be easily modified to implement other controls schemes in order to aid in the interpretation of AFM experiments.

  10. A simple and efficient quasi 3-dimensional viscoelastic model and software for simulation of tapping-mode atomic force microscopy

    PubMed Central

    2015-01-01

    Summary This paper introduces a quasi-3-dimensional (Q3D) viscoelastic model and software tool for use in atomic force microscopy (AFM) simulations. The model is based on a 2-dimensional array of standard linear solid (SLS) model elements. The well-known 1-dimensional SLS model is a textbook example in viscoelastic theory but is relatively new in AFM simulation. It is the simplest model that offers a qualitatively correct description of the most fundamental viscoelastic behaviors, namely stress relaxation and creep. However, this simple model does not reflect the correct curvature in the repulsive portion of the force curve, so its application in the quantitative interpretation of AFM experiments is relatively limited. In the proposed Q3D model the use of an array of SLS elements leads to force curves that have the typical upward curvature in the repulsive region, while still offering a very low computational cost. Furthermore, the use of a multidimensional model allows for the study of AFM tips having non-ideal geometries, which can be extremely useful in practice. Examples of typical force curves are provided for single- and multifrequency tapping-mode imaging, for both of which the force curves exhibit the expected features. Finally, a software tool to simulate amplitude and phase spectroscopy curves is provided, which can be easily modified to implement other controls schemes in order to aid in the interpretation of AFM experiments. PMID:26734515

  11. Statistics of reversible transitions in two-state trajectories in force-ramp spectroscopy.

    PubMed

    Diezemann, Gregor

    2014-05-14

    A possible way to extract information about the reversible dissociation of a molecular adhesion bond from force fluctuations observed in force ramp experiments is discussed. For small loading rates the system undergoes a limited number of unbinding and rebinding transitions observable in the so-called force versus extension (FE) curves. The statistics of these transient fluctuations can be utilized to estimate the parameters for the rebinding rate. This is relevant in the experimentally important situation where the direct observation of the reversed FE-curves is hampered, e.g., due to the presence of soft linkers. I generalize the stochastic theory of the kinetics in two-state models to the case of time-dependent kinetic rates and compute the relevant distributions of characteristic forces. While for irreversible systems there is an intrinsic relation between the rupture force distribution and the population of the free-energy well of the bound state, the situation is slightly more complex if reversible systems are considered. For a two-state model, a "stationary" rupture force distribution that is proportional to the population can be defined and allows to consistently discuss quantities averaged over the transient fluctuations. While irreversible systems are best analyzed in the soft spring limit of small pulling device stiffness and large loading rates, here I argue to use the stiffness of the pulling device as a control parameter in addition to the loading rate.

  12. Statistics of reversible transitions in two-state trajectories in force-ramp spectroscopy

    SciTech Connect

    Diezemann, Gregor

    2014-05-14

    A possible way to extract information about the reversible dissociation of a molecular adhesion bond from force fluctuations observed in force ramp experiments is discussed. For small loading rates the system undergoes a limited number of unbinding and rebinding transitions observable in the so-called force versus extension (FE) curves. The statistics of these transient fluctuations can be utilized to estimate the parameters for the rebinding rate. This is relevant in the experimentally important situation where the direct observation of the reversed FE-curves is hampered, e.g., due to the presence of soft linkers. I generalize the stochastic theory of the kinetics in two-state models to the case of time-dependent kinetic rates and compute the relevant distributions of characteristic forces. While for irreversible systems there is an intrinsic relation between the rupture force distribution and the population of the free-energy well of the bound state, the situation is slightly more complex if reversible systems are considered. For a two-state model, a “stationary” rupture force distribution that is proportional to the population can be defined and allows to consistently discuss quantities averaged over the transient fluctuations. While irreversible systems are best analyzed in the soft spring limit of small pulling device stiffness and large loading rates, here I argue to use the stiffness of the pulling device as a control parameter in addition to the loading rate.

  13. AFM nanoscale indentation in air of polymeric and hybrid materials with highly different stiffness

    NASA Astrophysics Data System (ADS)

    Suriano, Raffaella; Credi, Caterina; Levi, Marinella; Turri, Stefano

    2014-08-01

    In this study, nanomechanical properties of a variety of polymeric materials was investigated by means of AFM. In particular, selecting different AFM probes, poly(methyl methacrylate) (PMMA), polydimethylsiloxane (PDMS) bulk samples, sol-gel hybrid thin films and hydrated hyaluronic acid hydrogels were indented in air to determine the elastic modulus. The force-distance curves and the indentation data were found to be greatly affected by the cantilever stiffness and by tip geometry. AFM indentation tests show that the choice of the cantilever spring constant and of tip shape is crucially influenced by elastic properties of samples. When adhesion-dominated interactions occur between the tip and the surface of samples, force-displacement curves reveal that a suitable functionalization of AFM probes allows the control of such interactions and the extraction of Young' modulus from AFM curves that would be otherwise unfeasible. By applying different mathematical models depending on AFM probes and materials under investigation, the values of Young's modulus were obtained and compared to those measured by rheological and dynamic mechanical analysis or to literature data. Our results show that a wide range of elastic moduli (10 kPa-10 GPa) can be determined by AFM in good agreement with those measured by conventional macroscopic measurements.

  14. Probing Ternary Solvent Effect in High V(oc) Polymer Solar Cells Using Advanced AFM Techniques.

    PubMed

    Li, Chao; Ding, Yi; Soliman, Mikhael; Lorenzo, Josie; Dhasmana, Nitesh; Chantharasupawong, Panit; Ievlev, Anton V; Gesquiere, Andre J; Tetard, Laurene; Thomas, Jayan

    2016-02-01

    This work describes a simple method to develop a high V(oc) low band gap PSCs. In addition, two new atomic force microscopy (AFM)-based nanoscale characterization techniques to study the surface morphology and physical properties of the structured active layer are introduced. With the help of ternary solvent processing of the active layer and C60 buffer layer, a bulk heterojunction PSC with V(oc) more than 0.9 V and conversion efficiency 7.5% is developed. In order to understand the fundamental properties of the materials ruling the performance of the PSCs tested, AFM-based nanoscale characterization techniques including Pulsed-Force-Mode AFM (PFM-AFM) and Mode-Synthesizing AFM (MSAFM) are introduced. Interestingly, MSAFM exhibits high sensitivity for direct visualization of the donor-acceptor phases in the active layer of the PSCs. Finally, conductive-AFM (cAFM) studies reveal local variations in conductivity in the donor and acceptor phases as well as a significant increase in photocurrent in the PTB7:ICBA sample obtained with the ternary solvent processing. PMID:26807919

  15. Fabrication of a new substrate for atomic force microscopic observation of DNA molecules from an ultrasmooth sapphire plate.

    PubMed Central

    Yoshida, K; Yoshimoto, M; Sasaki, K; Ohnishi, T; Ushiki, T; Hitomi, J; Yamamoto, S; Sigeno, M

    1998-01-01

    A new stable substrate applicable to the observation of DNA molecules by atomic force microscopy (AFM) was fabricated from a ultrasmooth sapphire (alpha-Al2O3 single crystal) plate. The atomically ultrasmooth sapphire as obtained by high-temperature annealing has hydrophobic surfaces and could not be used for the AFM observation of DNA. However, sapphire treated with Na3PO4 aqueous solution exhibited a hydrophilic character while maintaining a smooth surface structure. The surface of the wet-treated sapphire was found by x-ray photoelectron spectroscopy and AFM to be approximately 0.3 nm. The hydrophilic surface character of the ultrasmooth sapphire plate made it easy for DNA molecules to adhere to the plate. Circular molecules of the plasmid DNA could be imaged by AFM on the hydrophilic ultrasmooth sapphire plate. PMID:9545030

  16. The dynamics and pH-dependence of Ag43 adhesins' self-association probed by atomic force spectroscopy

    NASA Astrophysics Data System (ADS)

    Jacquot, Adrien; Sakamoto, Chizuko; Razafitianamarahavo, Angelina; Caillet, Céline; Merlin, Jenny; Fahs, Ahmad; Ghigo, Jean-Marc; Duval, Jérôme F. L.; Beloin, Christophe; Francius, Grégory

    2014-10-01

    Self-associating auto-transporter (SAAT) adhesins are two-domain cell surface proteins involved in bacteria auto-aggregation and biofilm formation. Antigen 43 (Ag43) is a SAAT adhesin commonly found in Escherichia coli whose variant Ag43a has been shown to promote persistence of uropathogenic E. coli within the bladder. The recent resolution of the tri-dimensional structure of the 499 amino-acids' β-domain in Ag43a has shed light on the possible mechanism governing the self-recognition of SAAT adhesins, in particular the importance of trans-interactions between the L shaped β-helical scaffold of two α-domains of neighboring adhesins. In this study, we use single-molecule force spectroscopy (SMFS) and dynamic force spectroscopy (DFS) to unravel the dynamics of Ag43-self association under various pH and molecular elongation rate conditions that mimic the situations encountered by E. coli in its natural environment. Results evidenced an important stretchability of Ag43α with unfolding of sub-domains leading to molecular extension as long as 150 nm. Nanomechanical analysis of molecular stretching data suggested that self-association of Ag43 can lead to the formation of dimers and tetramers driven by rapid and weak cis- as well as slow but strong trans-interaction forces with a magnitude as large as 100-250 pN. The dynamics of cis- and trans-interactions were demonstrated to be strongly influenced by pH and applied shear force, thus suggesting that environmental conditions can modulate Ag43-mediated aggregation of bacteria at the molecular level.Self-associating auto-transporter (SAAT) adhesins are two-domain cell surface proteins involved in bacteria auto-aggregation and biofilm formation. Antigen 43 (Ag43) is a SAAT adhesin commonly found in Escherichia coli whose variant Ag43a has been shown to promote persistence of uropathogenic E. coli within the bladder. The recent resolution of the tri-dimensional structure of the 499 amino-acids' β-domain in Ag43a has shed

  17. Conformational rearrangements in the transmembrane domain of CNGA1 channels revealed by single-molecule force spectroscopy

    NASA Astrophysics Data System (ADS)

    Maity, Sourav; Mazzolini, Monica; Arcangeletti, Manuel; Valbuena, Alejandro; Fabris, Paolo; Lazzarino, Marco; Torre, Vincent

    2015-05-01

    Cyclic nucleotide-gated (CNG) channels are activated by binding of cyclic nucleotides. Although structural studies have identified the channel pore and selectivity filter, conformation changes associated with gating remain poorly understood. Here we combine single-molecule force spectroscopy (SMFS) with mutagenesis, bioinformatics and electrophysiology to study conformational changes associated with gating. By expressing functional channels with SMFS fingerprints in Xenopus laevis oocytes, we were able to investigate gating of CNGA1 in a physiological-like membrane. Force spectra determined that the S4 transmembrane domain is mechanically coupled to S5 in the closed state, but S3 in the open state. We also show there are multiple pathways for the unfolding of the transmembrane domains, probably caused by a different degree of α-helix folding. This approach demonstrates that CNG transmembrane domains have dynamic structure and establishes SMFS as a tool for probing conformational change in ion channels.

  18. Visualization of subsurface nanoparticles in a polymer matrix using resonance tracking atomic force acoustic microscopy and contact resonance spectroscopy.

    PubMed

    Kimura, Kuniko; Kobayashi, Kei; Yao, Atsushi; Yamada, Hirofumi

    2016-10-14

    A visualization technique of subsurface features with a nanometer-scale spatial resolution is strongly demanded. Some research groups have demonstrated the visualization of subsurface features using various techniques based on atomic force microscopy. However, the imaging mechanisms have not yet been fully understood. In this study, we demonstrated the visualization of subsurface Au nanoparticles buried in a polymer matrix 900 nm from the surface using two techniques; i.e., resonance tracking atomic force acoustic microscopy and contact resonance spectroscopy. It was clarified that the subsurface features were visualized by the two techniques as the area with a higher contact resonance frequency and a higher Q-factor than those in the surrounding area, which suggests that the visualization is realized by the variation of the contact stiffness and damping of the polymer matrix due to the existence of the buried nanoparticles. PMID:27607548

  19. Conformational rearrangements in the transmembrane domain of CNGA1 channels revealed by single-molecule force spectroscopy

    PubMed Central

    Maity, Sourav; Mazzolini, Monica; Arcangeletti, Manuel; Valbuena, Alejandro; Fabris, Paolo; Lazzarino, Marco; Torre, Vincent

    2015-01-01

    Cyclic nucleotide-gated (CNG) channels are activated by binding of cyclic nucleotides. Although structural studies have identified the channel pore and selectivity filter, conformation changes associated with gating remain poorly understood. Here we combine single-molecule force spectroscopy (SMFS) with mutagenesis, bioinformatics and electrophysiology to study conformational changes associated with gating. By expressing functional channels with SMFS fingerprints in Xenopus laevis oocytes, we were able to investigate gating of CNGA1 in a physiological-like membrane. Force spectra determined that the S4 transmembrane domain is mechanically coupled to S5 in the closed state, but S3 in the open state. We also show there are multiple pathways for the unfolding of the transmembrane domains, probably caused by a different degree of α-helix folding. This approach demonstrates that CNG transmembrane domains have dynamic structure and establishes SMFS as a tool for probing conformational change in ion channels. PMID:25963832

  20. Visualization of subsurface nanoparticles in a polymer matrix using resonance tracking atomic force acoustic microscopy and contact resonance spectroscopy.

    PubMed

    Kimura, Kuniko; Kobayashi, Kei; Yao, Atsushi; Yamada, Hirofumi

    2016-10-14

    A visualization technique of subsurface features with a nanometer-scale spatial resolution is strongly demanded. Some research groups have demonstrated the visualization of subsurface features using various techniques based on atomic force microscopy. However, the imaging mechanisms have not yet been fully understood. In this study, we demonstrated the visualization of subsurface Au nanoparticles buried in a polymer matrix 900 nm from the surface using two techniques; i.e., resonance tracking atomic force acoustic microscopy and contact resonance spectroscopy. It was clarified that the subsurface features were visualized by the two techniques as the area with a higher contact resonance frequency and a higher Q-factor than those in the surrounding area, which suggests that the visualization is realized by the variation of the contact stiffness and damping of the polymer matrix due to the existence of the buried nanoparticles.

  1. Visualization of subsurface nanoparticles in a polymer matrix using resonance tracking atomic force acoustic microscopy and contact resonance spectroscopy

    NASA Astrophysics Data System (ADS)

    Kimura, Kuniko; Kobayashi, Kei; Yao, Atsushi; Yamada, Hirofumi

    2016-10-01

    A visualization technique of subsurface features with a nanometer-scale spatial resolution is strongly demanded. Some research groups have demonstrated the visualization of subsurface features using various techniques based on atomic force microscopy. However, the imaging mechanisms have not yet been fully understood. In this study, we demonstrated the visualization of subsurface Au nanoparticles buried in a polymer matrix 900 nm from the surface using two techniques; i.e., resonance tracking atomic force acoustic microscopy and contact resonance spectroscopy. It was clarified that the subsurface features were visualized by the two techniques as the area with a higher contact resonance frequency and a higher Q-factor than those in the surrounding area, which suggests that the visualization is realized by the variation of the contact stiffness and damping of the polymer matrix due to the existence of the buried nanoparticles.

  2. An Atomic Force Microscopy based investigation of specific biomechanical properties for various types of neuronal cells

    NASA Astrophysics Data System (ADS)

    Spedden, Elise; White, James; Kaplan, David; Staii, Cristian

    2012-02-01

    Here we describe the use of Atomic Force Microscope (AFM) based techniques to characterize and explore the influence of biochemical and biomechanical cues on the growth and interaction of neuronal cells with surrounding guidance factors. Specifically, we use AFM topography and AFM force spectroscopy measurements to systematically investigate the morphology, elasticity, and real time growth of neuronal processes in the presence of different types of extracellular matrix proteins and growth factors. We therefore create a series of systems containing specified neuron densities where the type of the underlying growth promoting protein is different from sample to sample. For each system we measure key biomechanical parameters related to neuronal growth such as height and elastic modulus at multiple growth points on several types of neurons. We show that systematic measurements of these parameters yield fundamental information about the role played by substrate-plated guidance factors in determining elastic and morphological properties of neurons during growth.

  3. The influence of aminophylline on the nanostructure and nanomechanics of T lymphocytes: an AFM study.

    PubMed

    Huang, Xun; He, Jiexiang; Liu, Mingxian; Zhou, Changren

    2014-01-01

    Although much progress has been made in the illustration of the mechanism of aminophylline (AM) treating asthma, there is no data about its effect on the nanostructure and nanomechanics of T lymphocytes. Here, we presented atomic force spectroscopy (AFM)-based investigations at the nanoscale level to address the above fundamental biophysical questions. As increasing AM treatment time, T lymphocytes' volume nearly double increased and then decreased. The changes of nanostructural features of the cell membrane, i.e., mean height of particles, root-mean-square roughness (Rq), crack and fragment appearance, increased with AM treatment time. T lymphocytes were completely destroyed with 96-h treatment, and they existed in the form of small fragments. Analysis of force-distance curves showed that the adhesion force of cell surface decreased significantly with the increase of AM treatment time, while the cell stiffness increased firstly and then decreased. These changes were closely correlated to the characteristics and process of cell oncosis. In total, these quantitative and qualitative changes of T lymphocytes' structure and nanomechanical properties suggested that AM could induce T lymphocyte oncosis to exert anti-inflammatory effects for treating asthma. These findings provide new insights into the T lymphocyte oncosis and the anti-inflammatory mechanism and immune regulation actions of AM. PMID:25258618

  4. The influence of aminophylline on the nanostructure and nanomechanics of T lymphocytes: an AFM study

    NASA Astrophysics Data System (ADS)

    Huang, Xun; He, Jiexiang; Liu, Mingxian; Zhou, Changren

    2014-09-01

    Although much progress has been made in the illustration of the mechanism of aminophylline (AM) treating asthma, there is no data about its effect on the nanostructure and nanomechanics of T lymphocytes. Here, we presented atomic force spectroscopy (AFM)-based investigations at the nanoscale level to address the above fundamental biophysical questions. As increasing AM treatment time, T lymphocytes' volume nearly double increased and then decreased. The changes of nanostructural features of the cell membrane, i.e., mean height of particles, root-mean-square roughness (Rq), crack and fragment appearance, increased with AM treatment time. T lymphocytes were completely destroyed with 96-h treatment, and they existed in the form of small fragments. Analysis of force-distance curves showed that the adhesion force of cell surface decreased significantly with the increase of AM treatment time, while the cell stiffness increased firstly and then decreased. These changes were closely correlated to the characteristics and process of cell oncosis. In total, these quantitative and qualitative changes of T lymphocytes' structure and nanomechanical properties suggested that AM could induce T lymphocyte oncosis to exert anti-inflammatory effects for treating asthma. These findings provide new insights into the T lymphocyte oncosis and the anti-inflammatory mechanism and immune regulation actions of AM.

  5. Nanoscale rippling on polymer surfaces induced by AFM manipulation

    PubMed Central

    2015-01-01

    Summary Nanoscale rippling induced by an atomic force microscope (AFM) tip can be observed after performing one or many scans over the same area on a range of materials, namely ionic salts, metals, and semiconductors. However, it is for the case of polymer films that this phenomenon has been widely explored and studied. Due to the possibility of varying and controlling various parameters, this phenomenon has recently gained a great interest for some technological applications. The advent of AFM cantilevers with integrated heaters has promoted further advances in the field. An alternative method to heating up the tip is based on solvent-assisted viscoplastic deformations, where the ripples develop upon the application of a relatively low force to a solvent-rich film. An ensemble of AFM-based procedures can thus produce nanoripples on polymeric surfaces quickly, efficiently, and with an unprecedented order and control. However, even if nanorippling has been observed in various distinct modes and many theoretical models have been since proposed, a full understanding of this phenomenon is still far from being achieved. This review aims at summarizing the current state of the art in the perspective of achieving control over the rippling process on polymers at a nanoscale level. PMID:26733086

  6. Influence of atomic tip structure on the intensity of inelastic tunneling spectroscopy data analyzed by combined scanning tunneling spectroscopy, force microscopy, and density functional theory

    NASA Astrophysics Data System (ADS)

    Okabayashi, Norio; Gustafsson, Alexander; Peronio, Angelo; Paulsson, Magnus; Arai, Toyoko; Giessibl, Franz J.

    2016-04-01

    Achieving a high intensity in inelastic scanning tunneling spectroscopy (IETS) is important for precise measurements. The intensity of the IETS signal can vary by up to a factor of 3 for various tips without an apparent reason accessible by scanning tunneling microscopy (STM) alone. Here, we show that combining STM and IETS with atomic force microscopy enables carbon monoxide front-atom identification, revealing that high IETS intensities for CO/Cu(111) are obtained for single-atom tips, while the intensity drops sharply for multiatom tips. Adsorption of the CO molecule on a Cu adatom [CO/Cu/Cu(111)] such that the molecule is elevated over the substrate strongly diminishes the tip dependence of IETS intensity, showing that an elevated position channels most of the tunneling current through the CO molecule even for multiatom tips, while a large fraction of the tunneling current bypasses the CO molecule in the case of CO/Cu(111).

  7. Unraveling the Secrets of Bacterial Adhesion Organelles Using Single-Molecule Force Spectroscopy

    NASA Astrophysics Data System (ADS)

    Axner, Ove; Björnham, Oscar; Castelain, Mickaël; Koutris, Efstratios; Schedin, Staffan; Fällman, Erik; Andersson, Magnus

    Many types of bacterium express micrometer-long attachment organelles (so-called pili) whose role is to mediate adhesion to host tissue. Until recently, little was known about their function in the adhesion process. Force-measuring optical tweezers (FMOT) have since then been used to unravel the biomechanical properties of various types of pili, primarily those from uropathogenic E. coli, in particular their force-vs.-elongation response, but lately also some properties of the adhesin are situated at the distal end of the pilus. This knowledge provides an understanding of how piliated bacteria can sustain external shear forces caused by rinsing processes, e.g., urine flow. It has been found that many types of pilus exhibit unique and complex force-vs.-elongation responses. It has been conjectured that their dissimilar properties impose significant differences in their ability to sustain external forces and that different types of pilus therefore have dissimilar predisposition to withstand different types of rinsing conditions. An understanding of these properties is of high importance since it can serve as a basis for finding new means to combat bacterial adhesion, including that caused by antibiotic-resistance bacteria. This work presents a review of the current status of the assessment of biophysical properties of individual pili on single bacteria exposed to strain/stress, primarily by the FMOT technique. It also addresses, for the first time, how the elongation and retraction properties of the rod couple to the adhesive properties of the tip adhesin.

  8. Analysis of adhesive binding forces between laminin-1 and C2C12 muscle cell membranes measured via high resolution force spectroscopy

    NASA Astrophysics Data System (ADS)

    Gluck, George; Gilbert, Richard; Ortiz, Christine

    2002-03-01

    Laminins are a family of glycoproteins that regulate cell differentiation, shape, and motility through interactions with various cell surface receptors. Here, we have directly measured the biomolecular adhesive binding forces between a cantilever / probe tip that was covalently attached with laminin-1 and membrane receptors on C2C12 muscle cells using the technique of high-resolution force spectroscopy (HRFS). On retraction of the probe tip away from the membrane surface, discrete, long-range adhesive unbinding events were always observed. Statistical analysis of the data revealed an initial broad distribution of heterogeneous unbinding events (occurring at separation distances, D=0-2µm from the point of maximum compression) of magnitude 92.23±37.87pN followed by a narrow distribution of homogeneous unbinding events (occurring at D > 2µm) of magnitude 38.16±9.10pN, which is suggestive of an individual biomolecular adhesive interaction. On-going studies include loading rate dependence and effect of dystroglycan mutation.

  9. Co-axial Electrospun Polyacrylonitrile-Poly(methylmethacrylate) Nanofibers: Atomic Force Microscopy and Compositional Characterization

    PubMed Central

    Zander, N.E.; Strawhecker, K.E.; Orlicki, J.A.; Rawlett, A.M.; Beebe, T.P.

    2011-01-01

    Poly(methylmethacrylate) (PMMA)- Polyacrylonitrile (PAN) fibers were prepared using a conventional single-nozzle electrospinning technique. The as-spun fibers exhibited core-shell morphology as verified by transmission electron microscopy (TEM) and atomic force microscopy (AFM). AFM-phase and modulus mapping images of the fiber cross-section and x-ray photoelectron spectroscopy (XPS) analysis indicated PAN formed the shell and PMMA the core material. XPS, thermal gravimetric analysis (TGA), and elemental analysis were used to determine fiber compositional information. Soaking the fibers in solvent demonstrated removal of the core material, generating hollow PAN fibers. PMID:21928836

  10. Simultaneous AFM nano-patterning and imaging for photomask repair

    NASA Astrophysics Data System (ADS)

    Keyvani, Aliasghar; Tamer, Mehmet S.; van Es, Maarten H.; Sadeghian, Hamed

    2016-03-01

    In this paper we present a new AFM based nano-patterning technique that can be used for fast defect repairing of high resolution photomasks and possibly other high-speed nano-patterning applications. The proposed method works based on hammering the sample with tapping mode AFM followed by wet cleaning of the residuals. On the area where a specific pattern should be written, the tip-sample interaction force is tuned in a controlled manner by changing the excitation frequency of the cantilever without interrupting the imaging process. Using this method several patterns where transferred to different samples with imaging speed. While the pattern was transferred to the sample in each tracing scan line, the patterned sample was imaged in retracing scan line, thus the outcome was immediately visible during the experiment.

  11. FACS-sorted particles reduce the data variance in optical tweezers-assisted dynamic force spectroscopy measurements

    NASA Astrophysics Data System (ADS)

    Stangner, T.; Singer, D.; Wagner, C.; Gutsche, C.; Ueberschär, O.; Hoffmann, R.; Kremer, F.

    2013-08-01

    By combining optical tweezers-assisted dynamic force spectroscopy experiments with fluorescence activated cell sorting (FACS), we demonstrate a new approach to reducing the data variance in measuring receptor-ligand interactions on a single molecule level by ensuring similar coating densities. Therefore, the carboxyfluorescein-labelled monophosphorylated peptide tau226-240[pThr231] is anchored on melamine resin beads and these beads are sorted by FACS to achieve a homogeneous surface coverage. To quantify the impact of the fluorescence dye on the bond parameters between the phosphorylated peptide and the corresponding phosphorylation specific anti-human tau monoclonal antibody HPT-104, we perform dynamic force spectroscopy and compare the results to data using unsorted beads covered with the non-fluorescence peptide analogue. Finally, we demonstrate that the data variance of the relative binding frequency is significantly decreased by a factor of 3.4 using pre-sorted colloids with a homogeneous ligand coating compared to using unsorted colloids.

  12. Atomic force microscope with combined FTIR-Raman spectroscopy having a micro thermal analyzer

    DOEpatents

    Fink, Samuel D.; Fondeur, Fernando F.

    2011-10-18

    An atomic force microscope is provided that includes a micro thermal analyzer with a tip. The micro thermal analyzer is configured for obtaining topographical data from a sample. A raman spectrometer is included and is configured for use in obtaining chemical data from the sample.

  13. Ultrafast force-clamp spectroscopy to probe lac repressor-DNA interactions

    NASA Astrophysics Data System (ADS)

    Monico, Carina; Capitanio, Marco; Belcastro, Gionata; Vanzi, Francesco; Pavone, Francesco S.

    2013-06-01

    We recently developed an ultrafast force-clamp laser trap capable to probe, under controlled force, bimolecular interactions with unprecedented temporal resolution. Here we present the technique in the framework of protein-DNA interactions, specifically on Lactose repressor protein (LacI). The high temporal resolution of the method reveals the kinetics of both short- and long-lived interactions of LacI along the DNA template (from ˜100 μs to tens of seconds), as well the dependence on force of such interaction kinetics. The two kinetically well-distinct populations of interactions observed clearly represent specific interactions with the operator sequences and a fast scanning of LacI along non-cognate DNA. These results demonstrate the effectiveness of the method to study the sequence-dependent affinity of DNA-binding proteins along the DNA and the effects of force on a wide range of interaction durations, including μs time scales not accessible to other single-molecule methods. This improvement in time resolution provides also important means of investigation on the long-puzzled mechanism of target search on DNA and possible protein conformational changes occurring upon target recognition.

  14. Quantitative analysis of single-molecule force spectroscopy on folded chromatin fibers.

    PubMed

    Meng, He; Andresen, Kurt; van Noort, John

    2015-04-20

    Single-molecule techniques allow for picoNewton manipulation and nanometer accuracy measurements of single chromatin fibers. However, the complexity of the data, the heterogeneity of the composition of individual fibers and the relatively large fluctuations in extension of the fibers complicate a structural interpretation of such force-extension curves. Here we introduce a statistical mechanics model that quantitatively describes the extension of individual fibers in response to force on a per nucleosome basis. Four nucleosome conformations can be distinguished when pulling a chromatin fiber apart. A novel, transient conformation is introduced that coexists with single wrapped nucleosomes between 3 and 7 pN. Comparison of force-extension curves between single nucleosomes and chromatin fibers shows that embedding nucleosomes in a fiber stabilizes the nucleosome by 10 kBT. Chromatin fibers with 20- and 50-bp linker DNA follow a different unfolding pathway. These results have implications for accessibility of DNA in fully folded and partially unwrapped chromatin fibers and are vital for understanding force unfolding experiments on nucleosome arrays. PMID:25779043

  15. Quantitative analysis of single-molecule force spectroscopy on folded chromatin fibers

    PubMed Central

    Meng, He; Andresen, Kurt; van Noort, John

    2015-01-01

    Single-molecule techniques allow for picoNewton manipulation and nanometer accuracy measurements of single chromatin fibers. However, the complexity of the data, the heterogeneity of the composition of individual fibers and the relatively large fluctuations in extension of the fibers complicate a structural interpretation of such force-extension curves. Here we introduce a statistical mechanics model that quantitatively describes the extension of individual fibers in response to force on a per nucleosome basis. Four nucleosome conformations can be distinguished when pulling a chromatin fiber apart. A novel, transient conformation is introduced that coexists with single wrapped nucleosomes between 3 and 7 pN. Comparison of force-extension curves between single nucleosomes and chromatin fibers shows that embedding nucleosomes in a fiber stabilizes the nucleosome by 10 kBT. Chromatin fibers with 20- and 50-bp linker DNA follow a different unfolding pathway. These results have implications for accessibility of DNA in fully folded and partially unwrapped chromatin fibers and are vital for understanding force unfolding experiments on nucleosome arrays. PMID:25779043

  16. Oxide nanocrystal based nanocomposites for fabricating photoplastic AFM probes

    NASA Astrophysics Data System (ADS)

    Ingrosso, Chiara; Martin-Olmos, Cristina; Llobera, Andreu; Innocenti, Claudia; Sangregorio, Claudio; Striccoli, Marinella; Agostiano, Angela; Voigt, Anja; Gruetzner, Gabi; Brugger, Jürgen; Perez-Murano, Francesc; Curri, Maria Lucia

    2011-11-01

    We report on the synthesis, characterization and application of a novel nanocomposite made of a negative tone epoxy based photoresist modified with organic-capped Fe2O3 nanocrystals (NCs). The mechanical properties of the nanocomposite drastically improve upon incorporation of a suitable concentration of NCs in the polymer, without deteriorating its photolithography performance. High aspect ratio 3D microstructures made of the nanocomposite have been fabricated with a uniform surface morphology and with a resolution down to few micrometres. The embedded organic-capped Fe2O3 NCs drastically increase the stiffness and hardness of the epoxy based photoresist matrix, making the final material extremely interesting for manufacturing miniaturized polymer based mechanical devices and systems. In particular, the nanocomposite has been used as structural material for fabricating photoplastic Atomic Force Microscopy (AFM) probes with integrated tips showing outstanding mechanical response and high resolution imaging performance. The fabricated probes consist of straight cantilevers with low stress-gradient and high quality factors, incorporating sharp polymeric tips. They present considerably improved performance compared to pure epoxy based photoresist AFM probes, and to commercial silicon AFM probes.We report on the synthesis, characterization and application of a novel nanocomposite made of a negative tone epoxy based photoresist modified with organic-capped Fe2O3 nanocrystals (NCs). The mechanical properties of the nanocomposite drastically improve upon incorporation of a suitable concentration of NCs in the polymer, without deteriorating its photolithography performance. High aspect ratio 3D microstructures made of the nanocomposite have been fabricated with a uniform surface morphology and with a resolution down to few micrometres. The embedded organic-capped Fe2O3 NCs drastically increase the stiffness and hardness of the epoxy based photoresist matrix, making the

  17. Automated force controller for amplitude modulation atomic force microscopy.

    PubMed

    Miyagi, Atsushi; Scheuring, Simon

    2016-05-01

    Atomic Force Microscopy (AFM) is widely used in physics, chemistry, and biology to analyze the topography of a sample at nanometer resolution. Controlling precisely the force applied by the AFM tip to the sample is a prerequisite for faithful and reproducible imaging. In amplitude modulation (oscillating) mode AFM, the applied force depends on the free and the setpoint amplitudes of the cantilever oscillation. Therefore, for keeping the applied force constant, not only the setpoint amplitude but also the free amplitude must be kept constant. While the AFM user defines the setpoint amplitude, the free amplitude is typically subject to uncontrollable drift, and hence, unfortunately, the real applied force is permanently drifting during an experiment. This is particularly harmful in biological sciences where increased force destroys the soft biological matter. Here, we have developed a strategy and an electronic circuit that analyzes permanently the free amplitude of oscillation and readjusts the excitation to maintain the free amplitude constant. As a consequence, the real applied force is permanently and automatically controlled with picoNewton precision. With this circuit associated to a high-speed AFM, we illustrate the power of the development through imaging over long-duration and at various forces. The development is applicable for all AFMs and will widen the applicability of AFM to a larger range of samples and to a larger range of (non-specialist) users. Furthermore, from controlled force imaging experiments, the interaction strength between biomolecules can be analyzed. PMID:27250433

  18. Imaging spectroscopy of albedo and radiative forcing by light-absorbing impurities in mountain snow

    NASA Astrophysics Data System (ADS)

    Painter, Thomas H.; Seidel, Felix C.; Bryant, Ann C.; McKenzie Skiles, S.; Rittger, Karl

    2013-09-01

    Recent studies show that deposition of dust and black carbon to snow and ice accelerates snowmelt and perturbs regional climate and hydrologic cycles. Radiative forcing by aerosols is often neglected in climate and hydrological models in part due to scarcity of observations. Here we describe and validate an algorithm suite (Imaging Spectrometer-Snow Albedo and Radiative Forcing (IS-SnARF)) that provides quantitative retrievals of snow grain size, snow albedo, and radiative forcing by light-absorbing impurities in snow and ice (LAISI) from Airborne Visible/Infrared Imaging Spectrometer (AVIRIS) data collected on 15 June 2011 in the Senator Beck Basin Study Area (SBBSA), SW Colorado, USA. Radiative forcing by LAISI is retrieved by the integral of the convolution of spectral irradiance with spectral differences between the spectral albedo (scaled from the observed hemispherical-directional reflectance factor (HDRF)) and modeled clean snow spectral albedo. The modeled surface irradiance at time of acquisition at test sites was 1052 W m-2 compared to 1048 W m-2 measured with the field spectroradiometer measurements, a relative difference of 0.4%. HDRF retrievals at snow and bare soil sites had mean errors relative to in situ measurements of -0.4 ± 0.1% reflectance averaged across the spectrum and root-mean-square errors of 1.5 ± 0.1%. Comparisons of snow albedo and radiative forcing retrievals from AVIRIS with in situ measurements in SBBSA showed errors of 0.001-0.004 and 2.1 ± 5.1 W m-2, respectively. A counterintuitive result was that, in the presence of light absorbing impurities, near-surface snow grain size increased with elevation, whereas we generally expect that at lower elevation the grain size would be larger.

  19. Analysis of AFM cantilever dynamics close to sample surface

    NASA Astrophysics Data System (ADS)

    Habibnejad Korayem, A.; Habibnejad Korayem, Moharam; Ghaderi, Reza

    2013-07-01

    For imaging and manipulation of biological specimens application of atomic force microscopy (AFM) in liquid is necessary. In this paper, tapping-mode AFM cantilever dynamics in liquid close to sample surface is modeled and simulated by well defining the contact forces. The effect of cantilever tilting angle has been accounted carefully. Contact forces have some differences in liquid in comparison to air or vacuum in magnitude or formulation. Hydrodynamic forces are also applied on the cantilever due to the motion in liquid. A continuous beam model is used with its first mode and forward-time simulation method for simulation of its hybrid dynamics and the frequency response and amplitude versus separation diagrams are extracted. The simulation results show a good agreement with experimental results. The resonance frequency in liquid is so small in comparison to air due to additional mass and also additional damping due to the viscosity of the liquid around. The results show that the effect of separation on free vibration amplitude is great. Its effect on resonance frequency is considerable too.

  20. Electrostatic binding and hydrophobic collapse of peptide-nucleic acid aggregates quantified using force spectroscopy.

    PubMed

    Camunas-Soler, Joan; Frutos, Silvia; Bizarro, Cristiano V; de Lorenzo, Sara; Fuentes-Perez, Maria Eugenia; Ramsch, Roland; Vilchez, Susana; Solans, Conxita; Moreno-Herrero, Fernando; Albericio, Fernando; Eritja, Ramón; Giralt, Ernest; Dev, Sukhendu B; Ritort, Felix

    2013-06-25

    Knowledge of the mechanisms of interaction between self-aggregating peptides and nucleic acids or other polyanions is key to the understanding of many aggregation processes underlying several human diseases (e.g., Alzheimer's and Parkinson's diseases). Determining the affinity and kinetic steps of such interactions is challenging due to the competition between hydrophobic self-aggregating forces and electrostatic binding forces. Kahalalide F (KF) is an anticancer hydrophobic peptide that contains a single positive charge that confers strong aggregative properties with polyanions. This makes KF an ideal model to elucidate the mechanisms by which self-aggregation competes with binding to a strongly charged polyelectrolyte such as DNA. We use optical tweezers to apply mechanical forces to single DNA molecules and show that KF and DNA interact in a two-step kinetic process promoted by the electrostatic binding of DNA to the aggregate surface followed by the stabilization of the complex due to hydrophobic interactions. From the measured pulling curves we determine the spectrum of binding affinities, kinetic barriers, and lengths of DNA segments sequestered within the KF-DNA complex. We find there is a capture distance beyond which the complex collapses into compact aggregates stabilized by strong hydrophobic forces and discuss how the bending rigidity of the nucleic acid affects this process. We hypothesize that within an in vivo context, the enhanced electrostatic interaction of KF due to its aggregation might mediate the binding to other polyanions. The proposed methodology should be useful to quantitatively characterize other compounds or proteins in which the formation of aggregates is relevant. PMID:23706043

  1. Spectroscopy of quadrupole and octupole states in rare-earth nuclei from a Gogny force

    NASA Astrophysics Data System (ADS)

    Nomura, K.; Rodríguez-Guzmán, R.; Robledo, L. M.

    2015-07-01

    Collective quadrupole and octupole states are described in a series of Sm and Gd isotopes within the framework of the interacting boson model (IBM), whose Hamiltonian parameters are deduced from mean-field calculations with the Gogny energy density functional. The link between both frameworks is the (β2β3 ) potential energy surface computed within the Hartree-Fock-Bogoliubov framework in the case of the Gogny force. The diagonalization of the IBM Hamiltonian provides excitation energies and transition strengths of an assorted set of states including both positive- and negative-parity states. The resultant spectroscopic properties are compared with the available experimental data and also with the results of the configuration mixing calculations with the Gogny force within the generator coordinate method (GCM). The structure of excited 0+ states and its connection with double-octupole phonons is also addressed. The model is shown to describe the empirical trend of the low-energy quadrupole and octupole collective structure fairly well and turns out to be consistent with GCM results obtained with the Gogny force.

  2. Ultrastable Atomic Force Microscopy for Biophysics

    NASA Astrophysics Data System (ADS)

    Churnside, Allison B.

    Atomic force microscopy (AFM) is a multifunctional workhorse of nanoscience and molecular biophysics, but instrumental drift remains a critical issue that limits the precision and duration of experiments. We have significantly reduced the two most important types of drift: in position and in force. The first, position drift, is defined as uncontrolled motion between the tip and the sample, which occurs in all three dimensions. By scattering a laser off the apex of a commercial AFM tip, we locally measured and thereby actively controlled its three-dimensional position above a sample surface to <0.4 A (Deltaf = 0.01--10 Hz) in air at room temperature. With this enhanced stability, we demonstrated atomic-scale (˜1 A) tip-sample stability and registration over tens of minutes with a series of AFM images. The second type of drift is force drift. We found that the primary source of force drift for a popular class of soft cantilevers is their gold coating, even though they are coated on both sides to minimize drift. When the gold coating was removed through a simple chemical etch, this drift in deflection was reduced by more than an order of magnitude over the first 2 hours after wetting the tip. Removing the gold also led to ˜ 10-fold reduction in reflected light, yet short-term (0.1--10 s) force precision improved. With both position and force drift greatly reduced, the utility of the AFM is enhanced. These improvements led to several new AFM abilities, including a five-fold increase in the image signal-to-noise ratio; tip-registered, label-free optical imaging; registered tip return to a particular point on the sample; and dual-detection force spectroscopy, which enables a new extension clamp mode. We have applied these abilities to folding of both membrane and soluble proteins. In principle, the techniques we describe can be fully incorporated into many types of scanning probe microscopy, making this work a general improvement to scanning probe techniques.

  3. Spectroscopy of 26F to probe proton-neutron forces close to the drip line.

    PubMed

    Lepailleur, A; Sorlin, O; Caceres, L; Bastin, B; Borcea, C; Borcea, R; Brown, B A; Gaudefroy, L; Grévy, S; Grinyer, G F; Hagen, G; Hjorth-Jensen, M; Jansen, G R; Llidoo, O; Negoita, F; de Oliveira, F; Porquet, M-G; Rotaru, F; Saint-Laurent, M-G; Sohler, D; Stanoiu, M; Thomas, J C

    2013-02-22

    A long-lived J(π) = 4(1)(+) isomer, T(1/2) = 2.2(1) ms, has been discovered at 643.4(1) keV in the weakly bound (9)(26)F nucleus. It was populated at Grand Accélérateur National d'Ions Lourds in the fragmentation of a (36)S beam. It decays by an internal transition to the J(π) = 1(1)(+) ground state [82(14)%], by β decay to (26)Ne, or β-delayed neutron emission to (25)Ne. From the β-decay studies of the J(π) =1(1)(+) and J(π) = 4(1)(+) states, new excited states have been discovered in (25,26)Ne. Gathering the measured binding energies of the J(π) = 1(1)(+) -4(1)(+) multiplet in (9)(26)F, we find that the proton-neutron π0d(5/2)ν0d(3/2) effective force used in shell-model calculations should be reduced to properly account for the weak binding of (9)(26)F. Microscopic coupled cluster theory calculations using interactions derived from chiral effective field theory are in very good agreement with the energy of the low-lying 1(1)(+), 2(1)(+), 4(1)(+) states in (26)F. Including three-body forces and coupling to the continuum effects improve the agreement between experiment and theory as compared to the use of two-body forces only. PMID:23473138

  4. Analysis of the effect of LRP-1 silencing on the invasive potential of cancer cells by nanomechanical probing and adhesion force measurements using atomic force microscopy.

    PubMed

    Le Cigne, A; Chièze, L; Beaussart, A; El-Kirat-Chatel, S; Dufrêne, Y F; Dedieu, S; Schneider, C; Martiny, L; Devy, J; Molinari, M

    2016-04-01

    Low-density lipoprotein receptor-related protein 1 (LRP-1) can internalize proteases involved in cancer progression and is thus considered a promising therapeutic target. However, it has been demonstrated that LRP-1 is also able to regulate the endocytosis of membrane-anchored proteins. Thus, strategies that target LRP-1 to modulate proteolysis could also affect adhesion and cytoskeleton dynamics. Here, we investigated the effect of LRP-1 silencing on parameters reflecting cancer cells' invasiveness by atomic force microscopy (AFM). The results show that LRP-1 silencing induces changes in the cells' adhesion behavior, particularly the dynamics of cell attachment. Clear alterations in morphology, such as more pronounced stress fibers and increased spreading, leading to increased area and circularity, were also observed. The determination of the cells' mechanical properties by AFM showed that these differences are correlated with an increase in Young's modulus. Moreover, the measurements show an overall decrease in cell motility and modifications of directional persistence. An overall increase in the adhesion force between the LRP-1-silenced cells and a gelatin-coated bead was also observed. Ultimately, our AFM-based force spectroscopy data, recorded using an antibody directed against the β1 integrin subunit, provide evidence that LRP-1 silencing modifies the rupture force distribution. Together, our results show that techniques traditionally used for the investigation of cancer cells can be coupled with AFM to gain access to complementary phenotypic parameters that can help discriminate between specific phenotypes associated with different degrees of invasiveness. PMID:26965453

  5. Review and perspectives of AFM application on the study of deformable drop/bubble interactions.

    PubMed

    Wang, Wei; Li, Kai; Ma, Mengyu; Jin, Hang; Angeli, Panagiota; Gong, Jing

    2015-11-01

    The applications of Atomic Force Microscopy (AFM) on the study of dynamic interactions and film drainage between deformable bodies dispersed in aqueous solutions are reviewed in this article. Novel experimental designs and recent advances in experimental methodologies are presented, which show the advantage of using AFM as a tool for probing colloidal interactions. The effects of both DLVO and non-DLVO forces on the colloid stabilization mechanism are discussed. Good agreement is found between the force - drop/bubble deformation behaviour revealed by AFM measurements and the theoretical modeling of film drainage process, giving a convincing explanation of the occurrence of certain phenomenon. However, the behaviour and shape of deformable drops as they approach or retract is still not well resolved. In addition, when surfactants are present further research is needed on the absorption of surfactant molecules into the interfaces, their mobility and the effects on interfacial film properties.

  6. Atomic force microscopy to study intermolecular forces and bonds associated with bacteria.

    PubMed

    Lower, Steven K

    2011-01-01

    Atomic force microscopy (AFM) operates on a very different principle than other forms of microscopy, such as optical microscopy or electron microscopy. The key component of an AFM is a cantilever that bends in response to forces that it experiences as it touches another surface. Forces as small as a few picoNewtons can be detected and probed with AFM. AFM has become very useful in biological sciences because it can be used on living cells that are immersed in water. AFM is particularly useful when the cantilever is modified with chemical groups (e.g. amine or carboxylic groups), small beads (e.g. glass or latex), or even a bacterium. This chapter describes how AFM can be used to measure forces and bonds between a bacterium and another surface. This paper also provides an example of the use of AFM on Staphylococcus aureus, a Gram-positive bacterium that is often associated with biofilms in humans.

  7. Insight into mechanics of AFM tip-based nanomachining: bending of cantilevers and machined grooves

    NASA Astrophysics Data System (ADS)

    Al-Musawi, R. S. J.; Brousseau, E. B.; Geng, Y.; Borodich, F. M.

    2016-09-01

    Atomic force microscope (AFM) tip-based nanomachining is currently the object of intense research investigations. Values of the load applied to the tip at the free end of the AFM cantilever probe used for nanomachining are always large enough to induce plastic deformation on the specimen surface contrary to the small load values used for the conventional contact mode AFM imaging. This study describes an important phenomenon specific for AFM nanomachining in the forward direction: under certain processing conditions, the deformed shape of the cantilever probe may change from a convex to a concave orientation. The phenomenon can principally change the depth and width of grooves machined, e.g. the grooves machined on a single crystal copper specimen may increase by 50% on average following such a change in the deformed shape of the cantilever. It is argued that this phenomenon can take place even when the AFM-based tool is operated in the so-called force-controlled mode. The study involves the refined theoretical analysis of cantilever probe bending, the analysis of experimental signals monitored during the backward and forward AFM tip-based machining and the inspection of the topography of produced grooves.

  8. Insight into mechanics of AFM tip-based nanomachining: bending of cantilevers and machined grooves.

    PubMed

    Al-Musawi, R S J; Brousseau, E B; Geng, Y; Borodich, F M

    2016-09-23

    Atomic force microscope (AFM) tip-based nanomachining is currently the object of intense research investigations. Values of the load applied to the tip at the free end of the AFM cantilever probe used for nanomachining are always large enough to induce plastic deformation on the specimen surface contrary to the small load values used for the conventional contact mode AFM imaging. This study describes an important phenomenon specific for AFM nanomachining in the forward direction: under certain processing conditions, the deformed shape of the cantilever probe may change from a convex to a concave orientation. The phenomenon can principally change the depth and width of grooves machined, e.g. the grooves machined on a single crystal copper specimen may increase by 50% on average following such a change in the deformed shape of the cantilever. It is argued that this phenomenon can take place even when the AFM-based tool is operated in the so-called force-controlled mode. The study involves the refined theoretical analysis of cantilever probe bending, the analysis of experimental signals monitored during the backward and forward AFM tip-based machining and the inspection of the topography of produced grooves. PMID:27532247

  9. Probing ternary solvent effect in high Voc polymer solar cells using advanced AFM techniques

    DOE PAGES

    Li, Chao; Soleman, Mikhael; Lorenzo, Josie; Dhasmana, Nitesh; Chantharasupawong, Panit; Ievlev, Anton; Gesquiere, Andre; Tetard, Laurene; Thomas, Jayan

    2016-01-25

    This work describes a simple method to develop a high Voc low band gap PSCs. In addition, two new atomic force microscopy (AFM)-based nanoscale characterization techniques to study the surface morphology and physical properties of the structured active layer are introduced. With the help of ternary solvent processing of the active layer and C60 buffer layer, a bulk heterojunction PSC with Voc more than 0.9 V and conversion efficiency 7.5% is developed. In order to understand the fundamental properties of the materials ruling the performance of the PSCs tested, AFM-based nanoscale characterization techniques including Pulsed-Force-Mode AFM (PFM-AFM) and Mode-Synthesizing AFMmore » (MSAFM) are introduced. Interestingly, MSAFM exhibits high sensitivity for direct visualization of the donor–acceptor phases in the active layer of the PSCs. Lastly, conductive-AFM (cAFM) studies reveal local variations in conductivity in the donor and acceptor phases as well as a significant increase in photocurrent in the PTB7:ICBA sample obtained with the ternary solvent processing.« less

  10. Non-specific binding of Na+ and Mg2+ to RNA determined by force spectroscopy methods.

    PubMed

    Bizarro, C V; Alemany, A; Ritort, F

    2012-08-01

    RNA duplex stability depends strongly on ionic conditions, and inside cells RNAs are exposed to both monovalent and multivalent ions. Despite recent advances, we do not have general methods to quantitatively account for the effects of monovalent and multivalent ions on RNA stability, and the thermodynamic parameters for secondary structure prediction have only been derived at 1M [Na(+)]. Here, by mechanically unfolding and folding a 20 bp RNA hairpin using optical tweezers, we study the RNA thermodynamics and kinetics at different monovalent and mixed monovalent/Mg(2+) salt conditions. We measure the unfolding and folding rupture forces and apply Kramers theory to extract accurate information about the hairpin free energy landscape under tension at a wide range of ionic conditions. We obtain non-specific corrections for the free energy of formation of the RNA hairpin and measure how the distance of the transition state to the folded state changes with force and ionic strength. We experimentally validate the Tightly Bound Ion model and obtain values for the persistence length of ssRNA. Finally, we test the approximate rule by which the non-specific binding affinity of divalent cations at a given concentration is equivalent to that of monovalent cations taken at 100-fold concentration for small molecular constructs. PMID:22492710

  11. Effect of water chemistry and aging on iron-mica interaction forces: implications for iron particle transport.

    PubMed

    Pensini, Erica; Yip, Christopher M; O'Carroll, Denis M; Sleep, Brent E

    2012-07-17

    The transport of particles through groundwater systems is governed by a complex interplay of mechanical and chemical forces that are ultimately responsible for binding to geological substrates. To understand these forces in the context of zero valent iron particles used in the remediation of groundwater, atomic force microscopy (AFM)-based force spectroscopy was employed to characterize the interactions between AFM tips modified with either carbonyl iron particles (CIP) or electrodeposited Fe as a function of counterion valency, temperature, particle morphology, and age. The measured interaction forces were always attractive for both fresh and aged CIP and electrodeposited iron, except in 100 mM NaCl, as a consequence of electrostatic attraction between the negatively charged mica and positively charged iron. In 100 mM NaCl, repulsive hydration forces appeared to dominate. Good agreement was found between the experimental data and predictions based on the extended DLVO (XDLVO) theory. The effect of aging on iron particle composition and morphology was assessed by X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) revealing that the aged particles comprising a zero valent iron core passivated by a mixture of iron oxides and hydroxides. Force spectroscopy showed that aging caused variations in the adhesive force due to the changes in particle morphology and contact area.

  12. Fabrication of cone-shaped boron doped diamond and gold nanoelectrodes for AFM-SECM

    NASA Astrophysics Data System (ADS)

    Avdic, A.; Lugstein, A.; Wu, M.; Gollas, B.; Pobelov, I.; Wandlowski, T.; Leonhardt, K.; Denuault, G.; Bertagnolli, E.

    2011-04-01

    We demonstrate a reliable microfabrication process for a combined atomic force microscopy (AFM) and scanning electrochemical microscopy (SECM) measurement tool. Integrated cone-shaped sensors with boron doped diamond (BDD) or gold (Au) electrodes were fabricated from commercially available AFM probes. The sensor formation process is based on mature semiconductor processing techniques, including focused ion beam (FIB) machining, and highly selective reactive ion etching (RIE). The fabrication approach preserves the geometry of the original AFM tips resulting in well reproducible nanoscaled sensors. The feasibility and functionality of the fully featured tips are demonstrated by cyclic voltammetry, showing good agreement between the measured and calculated currents of the cone-shaped AFM-SECM electrodes.

  13. Fabrication of cone-shaped boron doped diamond and gold nanoelectrodes for AFM-SECM.

    PubMed

    Avdic, A; Lugstein, A; Wu, M; Gollas, B; Pobelov, I; Wandlowski, T; Leonhardt, K; Denuault, G; Bertagnolli, E

    2011-04-01

    We demonstrate a reliable microfabrication process for a combined atomic force microscopy (AFM) and scanning electrochemical microscopy (SECM) measurement tool. Integrated cone-shaped sensors with boron doped diamond (BDD) or gold (Au) electrodes were fabricated from commercially available AFM probes. The sensor formation process is based on mature semiconductor processing techniques, including focused ion beam (FIB) machining, and highly selective reactive ion etching (RIE). The fabrication approach preserves the geometry of the original AFM tips resulting in well reproducible nanoscaled sensors. The feasibility and functionality of the fully featured tips are demonstrated by cyclic voltammetry, showing good agreement between the measured and calculated currents of the cone-shaped AFM-SECM electrodes.

  14. Fabrication of cone-shaped boron doped diamond and gold nanoelectrodes for AFM-SECM.

    PubMed

    Avdic, A; Lugstein, A; Wu, M; Gollas, B; Pobelov, I; Wandlowski, T; Leonhardt, K; Denuault, G; Bertagnolli, E

    2011-04-01

    We demonstrate a reliable microfabrication process for a combined atomic force microscopy (AFM) and scanning electrochemical microscopy (SECM) measurement tool. Integrated cone-shaped sensors with boron doped diamond (BDD) or gold (Au) electrodes were fabricated from commercially available AFM probes. The sensor formation process is based on mature semiconductor processing techniques, including focused ion beam (FIB) machining, and highly selective reactive ion etching (RIE). The fabrication approach preserves the geometry of the original AFM tips resulting in well reproducible nanoscaled sensors. The feasibility and functionality of the fully featured tips are demonstrated by cyclic voltammetry, showing good agreement between the measured and calculated currents of the cone-shaped AFM-SECM electrodes. PMID:21368355

  15. Spectral and AFM characterization of trimethylammoniophenylporphyrin and concanavalin A associate in solution and monolithic SiO 2 gels obtained by the sol-gel method

    NASA Astrophysics Data System (ADS)

    Polska, Katarzyna; Radzki, Stanisław

    2008-06-01

    The associate between water-soluble cationic tetrakis[4-(trimethylammonio)phenyl] porphyrin (H2TTMePP) and concanavalin A (Con A) has been studied in the tris-buffer solution by absorption and emission electron spectroscopy. The porphyrin and porphyrin concanavalin associate has been incorporated into the monolithic pure silica gels obtained by polycondensation of tetraethoxysilane. The optically transparent dried gels were studied using absorption and fluorescence spectroscopic techniques and also by the tapping mode of atomic force microscopy (AFM). Complex formation between porphyrin and concanavalin takes place in both solution and gel. In these media porphyrin and its lectin associate exhibit luminescence emission in the vis-ir range when excited with visible light. Upon binding to concanavalin A the increase in porphyrin fluorescence intensity and the red-shift in the absorption and emission maxima have been observed. AFM visualisation of porphyrin and the porphyrin-concanavalin conjugate shows significant differences between nanostructures of the pure porphyrin and complex doped gels. It has been found that the ''smooth'' surfaces of silica gels prepared by the sol-gel technique are an excellent medium for the AFM visualisation of biomolecules.

  16. LET Spectrum Measurements In CR-39 PNTD With AFM

    SciTech Connect

    Johnson, C. E.; DeWitt, J. M.; Benton, E. R.; Yasuda, N.; Benton, E. V.

    2011-06-01

    Energetic protons, neutrons, and heavy ions undergoing collisions with target nuclei of varying Z can produce residual heavy recoil fragments via intra-nuclear cascade/evaporation reactions. The particles produced in these non-elastic collisions generally have such extremely short range ({approx}<10 {mu}m) that they cannot be directly observed by conventional detection methods including CR-39 plastic nuclear track detector (PNTD) that has been chemically etched for analysis by standard visible light microscopy. However, high-LET recoil fragments having range on the order of several cell diameters can be produced in tissue during radiotherapy using proton and carbon beams. We have developed a method to analyze short-range, high-LET tracks in CR-39 plastic nuclear track detector (PNTD) using short duration chemical etching ({approx}<1 {mu}m) following by automated atomic force microscope (AFM) scanning. The post-scan data processing used in this work was based on semi-automated matrix analysis opposed to traditional grey-scale image analysis. This method takes advantage of the 3-D data obtained via AFM to achieve robust discrimination of nuclear tracks from other features inherently present in the post-etch detector surface. Through automation of AFM scanning, sufficient AFM scan frames were obtained to attain an LET spectrum spanning the LET range from 200-1500 keV/{mu}m. In addition to our experiments, simulations were carried out with the Monte Carlo transport code, FLUKA. To demonstrate this method, CR-39 PNTD was exposed to the proton therapy beam at Loma Linda University Medical Center (LLUMC) at 60 and 230 MeV. Additionally, detectors were exposed to 1 GeV protons at the NASA Space Radiation Laboratory (NSRL) at Brookhaven National Laboratory (BNL). For these exposures CR-39 PNTD, Al and Cu target foils were used between detector layers.

  17. LET Spectrum Measurements In CR-39 PNTD With AFM

    NASA Astrophysics Data System (ADS)

    Johnson, C. E.; DeWitt, J. M.; Benton, E. R.; Yasuda, N.; Benton, E. V.

    2011-06-01

    Energetic protons, neutrons, and heavy ions undergoing collisions with target nuclei of varying Z can produce residual heavy recoil fragments via intra-nuclear cascade/evaporation reactions. The particles produced in these non-elastic collisions generally have such extremely short range (˜<10 μm) that they cannot be directly observed by conventional detection methods including CR-39 plastic nuclear track detector (PNTD) that has been chemically etched for analysis by standard visible light microscopy. However, high-LET recoil fragments having range on the order of several cell diameters can be produced in tissue during radiotherapy using proton and carbon beams. We have developed a method to analyze short-range, high-LET tracks in CR-39 plastic nuclear track detector (PNTD) using short duration chemical etching (˜<1 μm) following by automated atomic force microscope (AFM) scanning. The post-scan data processing used in this work was based on semi-automated matrix analysis opposed to traditional grey-scale image analysis. This method takes advantage of the 3-D data obtained via AFM to achieve robust discrimination of nuclear tracks from other features inherently present in the post-etch detector surface. Through automation of AFM scanning, sufficient AFM scan frames were obtained to attain an LET spectrum spanning the LET range from 200-1500 keV/μm. In addition to our experiments, simulations were carried out with the Monte Carlo transport code, FLUKA. To demonstrate this method, CR-39 PNTD was exposed to the proton therapy beam at Loma Linda University Medical Center (LLUMC) at 60 and 230 MeV. Additionally, detectors were exposed to 1 GeV protons at the NASA Space Radiation Laboratory (NSRL) at Brookhaven National Laboratory (BNL). For these exposures CR-39 PNTD, Al and Cu target foils were used between detector layers.

  18. Quantitative nano-mechanics of biological cells with AFM

    NASA Astrophysics Data System (ADS)

    Sokolov, Igor

    2013-03-01

    The importance of study of living cells is hard to overestimate. Cell mechanics is a relatively young, yet not a well-developed area. Besides just a fundamental interest, large practical need has emerged to measure cell mechanics quantitatively. Recent studies revealed a significant correlation between stiffness of biological cells and various human diseases, such as cancer, malaria, arthritis, and even aging. However, really quantitative studies of mechanics of biological cells are virtually absent. It is not even clear if the cell, being a complex and heterogeneous object, can be described by the elastic modulus at all. Atomic force microscopy (AFM) is a natural instrument to study properties of cells in their native environments. Here we will demonstrate that quantitative measurements of elastic modulus of cells with AFM are possible. Specifically, we will show that the ``cell body'' (cell without ``brush'' surface layer, a non-elastic layer surrounding cells) typically demonstrates the response of a homogeneous elastic medium up to the deformation of 10-20%, but if and only if a) the cellular brush layer is taken into account, b) rather dull AFM probes are used. This will be justified with the help of the strong condition of elastic behavior of material: the elastic modulus is shown to be independent on the indentation depth. We will also demonstrate that an attempt either to ignore the brush layer or to use sharp AFM probes will result in the violation of the strong condition, which implies impossibility to use the concept of the elastic modulus to describe cell mechanics in such experiments. Examples of quantitative measurements of the Young's modulus of the cell body and the cell brush parameters will be given for various cells. Address when submitting: Clarkson University, Potsdam, NY 13699

  19. LET spectrum measurements in Cr-39 PNTD with AFM

    SciTech Connect

    Johnson, Carl Edward; De Witt, Joel M; Benton, Eric R; Yasuda, Nakahiro; Benton, Eugene V

    2010-01-01

    Energetic protons, neutrons, and heavy ions undergoing collisions with target nuclei of varying Z can produce residual heavy recoil fragments via intra-nuclear cascade/evaporation reactions. The particles produced in these non-elastic collisions generally have such extremely short range ({approx}< 10 {mu}m) that they cannot be directly observed by conventional detection methods including CR-39 plastic nuclear track detector (PNTD) that has been chemically etched for analysis by standard visible light microscopy. However, high-LET recoil fragments having range on the order of several cell diameters can be produced in tissue during radiotherapy using proton and carbon beams. We have developed a method to analyze short-range, high-LET tracks in CR-39 plastic nuclear track detector (PNTD) using short duration chemical etching ({approx}< 1 {mu}m) followed by automated atomic force microscope (AFM) scanning. The post-scan data processing used in this work was based on semi-automated matrix analysis opposed to traditional grey-scale image analysis. This method takes advantage of the 3-D data obtained via AFM to achieve robust discrimination of nuclear tracks from other features. Through automation of AFM scanning, sufficient AFM scan frames were obtained to attain an LET spectrum spanning the LET range from 200-1500 keV/{mu}m. In addition to our experiments, simulations were carried out with the Monte Carlo transport code, FLUKA. To demonstrate this method, CR-39 PNTD was exposed to the proton therapy beam at Loma Linda University Medical Center (LLUMC) at 60 and 230 MeV. Additionally, detectors were exposed to I GeV protons at the NASA Space Radiation Laboratory (NSRL) at Brookhaven National Laboratory (BNL). For these exposures CR-39 PNTD, Al and Cu target foils were used between detector layers.

  20. Modeling AFM Induced Mechanical Deformation of Living Cells

    SciTech Connect

    Rudd, R E; McElfresh, M; Balhorn, R; Allen, M J; Belak, J

    2002-11-15

    Finite element modeling has been applied to study deformation of living cells in Atomic Force Microscopy (AFM) and particularly Recognition Force Microscopy (RFM). The abstract mechanical problem of interest is the response to RFM point loads of an incompressible medium enclosed in a fluid membrane. Cells are soft systems, susceptible to large deformations in the course of an RFM measurement. Often the local properties such as receptor anchoring forces, the reason for the measurement, are obscured by the response of the cell as a whole. Modeling can deconvolute these effects. This facilitates experimental efforts to have reproducible measurements of mechanical and chemical properties at specific kinds of receptor sites on the membrane of a living cell. In this article we briefly review the RFM technique for cells and the problems it poses, and then report on recent progress in modeling the deformation of cells by a point load.

  1. Viscoelasticity of gelatin surfaces probed by AFM noise analysis.

    PubMed

    Benmouna, Farida; Johannsmann, Diethelm

    2004-01-01

    The viscoelastic properties of surfaces of swollen gelatin were investigated by analyzing the Brownian motion of an atomic force microscopy (AFM) cantilever in contact with the gel surface. A micron-sized glass sphere attached to the AFM cantilever is used as the dynamic probe. When the sphere approaches the gelatin surface, there is a static repulsive force without a jump into contact. The cantilever's Brownian movement is monitored in parallel, providing access to the dynamic sphere-surface interaction as quantified by the dynamic spring constant, kappa, and the drag coefficient, xi. Gelatin is used as a model substance for a variety of other soft surfaces, where the stiffness of the gel can be varied via the solvent quality, the bloom number, and the pH. The modulus derived from the static force-distance curve is in the kPa range, consistent with the literature. However, the dynamic spring constant as derived from the Brownian motion is much larger than the static differential spring constant dF/dz. On retraction, one observes a rather strong adhesion hysteresis. The strength of the bridge (as given by the dynamic spring constant and the drag coefficient) is very small. PMID:15745019

  2. Depositions of molecular nanomagnets on graphene investigated with atomic force microscopy and Raman spectroscopy

    NASA Astrophysics Data System (ADS)

    Walker, Sean; Vojvodin, Cameron; Li, Zhi; Willick, Kyle; Tang, Xiaowu (Shirley); Baugh, Jonathan

    Molecular nanomagnets display interesting quantum phenomena, and have been proposed as potential building blocks in a variety of nanoelectronic devices with applications to both quantum memory and quantum information processing. These devices often require deposition of the molecules either sparsely (e.g. for single molecule devices) or as a thin-film. Consequently, in order for these devices to be successfully realized, the nature of the interactions between nanomagnets and the surfaces on which they may be deposited needs to be understood. We have investigated the depositions of molecular nanomagnets on graphene using atomic force microscopy and Raman spectrocopy. The nanomagnets contained a range of chemical functional groups including long alkyl chains and extended π-systems of electrons. By comparing their binding affinities we learn about the nature of the interactions between the different functional groups and the graphene.

  3. Adsorption Behavior of Cellulose and Its Derivatives toward Ag(I) in Aqueous Medium: An AFM, Spectroscopic, and DFT Study.

    PubMed

    Zhu, Chuantao; Dobryden, Illia; Rydén, Jens; Öberg, Sven; Holmgren, Allan; Mathew, Aji P

    2015-11-17

    The aim of this study was to develop a fundamental understanding of the adsorption behavior of metal ions on cellulose surfaces using experimental techniques supported by computational modeling, taking Ag(I) as an example. Force interactions among three types of cellulose microspheres (native cellulose and its derivatives with sulfate and phosphate groups) and the silica surface in AgNO3 solution were studied with atomic force microscopy (AFM) using the colloidal probe technique. The adhesion force between phosphate cellulose microspheres (PCM) and the silica surface in the aqueous AgNO3 medium increased significantly with increasing pH while the adhesion force slightly decreased for sulfate cellulose microspheres (SCM), and no clear adhesion force was observed for native cellulose microspheres (CM). The stronger adhesion enhancement for the PCM system is mainly attributed to the electrostatic attraction between Ag(I) and the negative silica surface. The observed force trends were in good agreement with the measured zeta potentials. The scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) analyses confirmed the presence of silver on the surface of cellulose microspheres after adsorption. This study showed that PCM with a high content of phosphate groups exhibited a larger amount of adsorbed Ag(I) than CM and SCM and possible clustering of Ag(I) to nanoparticles. The presence of the phosphate group and a wavenumber shift of the P-OH vibration caused by the adsorption of silver ions on the phosphate groups were further confirmed with computational studies using density functional theory (DFT), which gives support to the above findings regarding the adsorption and clustering of Ag(I) on the cellulose surface decorated with phosphate groups as well as IR spectra.

  4. Cell secretion machinery: studies using the AFM.

    PubMed

    Jena, Bhanu P

    2006-01-01

    A new field in biology, 'nano-cell biology', has emerged from the successful use of force microscopy in understanding the structure and dynamics of cells and biomolecules, at nm resolution and in real time. Atomic force microscopy, in combination with conventional tools and approaches (electron microscopy, electrophysiology, X-ray diffraction, photon correlation spectroscopy, mass spectroscopy, biochemistry, and molecular biology), has revealed for the first time, the universal molecular machinery and mechanism of secretion in cells. Secretion occurs in all living cells and involves the delivery of intracellular products to the cell exterior. Secretory products are packaged and stored in membranous sacs or vesicles within the cell. When the cell needs to secrete these products, the secretory vesicles containing them, dock and fuse at plasma membrane-associated supramolecular structures called Porosome, to release their contents. Specialized cells for neurotransmission, enzyme secretion, or hormone release utilize a highly regulated secretory process. During secretion, swelling of secretory vesicles results in a build-up of intravesicular pressure, allowing expulsion of vesicular contents. The extent of vesicle swelling dictates the amount of vesicular contents expelled. The discovery of the porosome as the universal secretory machinery, its isolation, its structure and dynamics at nm resolution and in real time, its biochemical composition and functional reconstitution into artificial lipid membrane, have been determined. The molecular mechanism of secretory vesicle swelling, and the fusion of opposing bilayers, i.e., the fusion of secretory vesicle membrane at the base of the porosome membrane, has also been resolved.

  5. Calibration of a dual-trap optical tweezers for single molecule force spectroscopy study

    NASA Astrophysics Data System (ADS)

    Wang, Guoqing; Hu, Chunguang; Gao, Xiaoqing; Su, Chenguang; Wang, Sirong; Lei, Hai; Hu, Xiaodong; Li, Hongbin; Hu, Xiaotang

    2015-10-01

    Optical tweezers has shown its significant advantages in applying pico-Newton force on micro beads and handling them with nanometer-level precision, and becomes a powerful tool for single-molecule biology. Many excellent researching results in use of the optical tweezers have been reported. Most of them focus on the single-trap optical tweezers experiments. However, when a single-trap optical tweezers is applied to biological molecule, there is often an obvious noise from the sample chamber holder to which one end of the sample molecule is tethered. In contrast, a dual-trap optical tweezers can intrinsically avoid this problem because both ends of the sample tethered to microspheres are manipulated with two separate optical traps. In order to force the molecule precisely, it is of importance to do calibrations for both traps. Many approaches have been studied to obtain the stiffness and sensitivity of the trap, but those are not quite suitable for making calibration during experiment. Here, we use a modified method of power spectrum density (PSD) for the calibrations of the stiffness and sensitivity of the traps, which combines a sinusoidal motion of the sample stage. The main strength of the method is that the beads used for the calibration also can be used in experiment later. In addition, the calibration can be performed during experiment. Finally, an experiment using a dsDNA molecule to test the system is presented. The results show that the calibration approach for the dual-trap optical tweezers is efficient and accurate.

  6. Germ direct observation by AFM under crystallization of self-organized assemblies of mono-protonated meso-tetraphenylporphine dimers

    NASA Astrophysics Data System (ADS)

    Udal'tsov, Alexander V.

    2016-08-01

    Assemblies consisting of mono-protonated meso-tetraphenylporphine dimers and water have been investigated by UV-vis spectroscopy in solution and atomic force microscopy (AFM) in thin layers. These assemblies self-organized into domains produce microcrystals in thin layer. Morphology of the microcrystals and characteristic features of crystallization germ on the top found by contact AFM indicate that surface tension of an aqueous layer on the domain generates the crystallization process. Estimations of the pressure producing the germ and bulk modulus (Bm) of microcrystals give 26.3±2.6 MPa and 3.72 GPa and Bm=12.7 GPa obtained for dried thin films. The former modulus is comparable with bulk modulus of water (2.174 GPa) that implies liquid crystals formation. Absorptions of longitudinal optical (LO) phonons with ћωLO=0.3761 and 0.3577 eV, which are arisen because of hole polaron moving through water, are found in the electronic spectra of the assemblies. The crystallization is suggested to occur due to Zundel cation (H5O2+) operation like the water-porphyrin matrix self-organization found earlier.

  7. Characterization of microscale wear in a ploysilicon-based MEMS device using AFM and PEEM-NEXAFS spectromicroscopy.

    SciTech Connect

    Grierson, D. S.; Konicek, A. R.; Wabiszewski, G. E.; Sumant, A. V.; de Boer, M. P.; Corwin, A. D.; Carpick, R. W.

    2009-12-01

    Mechanisms of microscale wear in silicon-based microelectromechanical systems (MEMS) are elucidated by studying a polysilicon nanotractor, a device specifically designed to conduct friction and wear tests under controlled conditions. Photoelectron emission microscopy (PEEM) was combined with near-edge X-ray absorption fine structure (NEXAFS) spectroscopy and atomic force microscopy (AFM) to quantitatively probe chemical changes and structural modification, respectively, in the wear track of the nanotractor. The ability of PEEM-NEXAFS to spatially map chemical variations in the near-surface region of samples at high lateral spatial resolution is unparalleled and therefore ideally suited for this study. The results show that it is possible to detect microscopic chemical changes using PEEM-NEXAFS, specifically, oxidation at the sliding interface of a MEMS device. We observe that wear induces oxidation of the polysilicon at the immediate contact interface, and the spectra are consistent with those from amorphous SiO{sub 2}. The oxidation is correlated with gouging and debris build-up in the wear track, as measured by AFM and scanning electron microscopy (SEM).

  8. Nonadditive intermolecular forces from the spectroscopy of van der Waals trimers: A theoretical study of Ar[sub 2]-HF

    SciTech Connect

    Ernesti, A.; Hutson, J.M. )

    1995-01-01

    Calculations of vibrational energies and rotational constants are carried out for the van der Waals trimer Ar[sub 2]-HF. The calculations include all five intermolecular degrees of freedom. The different intramolecular vibrational states [ital v] of the HF molecule are separated out adiabatically, so that the calculations are carried out on effective intermolecular potentials for each HF vibrational state separately. Calculations are performed both on pairwise-additive potentials, derived from the well-known Ar-Ar and Ar-HF potentials, and on nonadditive potentials, incorporating various different contributions to the three-body forces. The results are compared with experimental results from high-resolution spectroscopy, and provide detailed information on the anisotropy of the nonadditive intermolecular forces. As in previous work on Ar[sub 2]-HCl, it is found that a very important nonadditive term arises from the interaction between the permanent multipoles of the HF molecule and the exchange quadrupole caused by distortion of the two Ar atoms as they overlap. An improved model of this term is described.

  9. Rapid identification of bacteria utilizing amplified dielectrophoretic force-assisted nanoparticle-induced surface-enhanced Raman spectroscopy

    PubMed Central

    2014-01-01

    Dielectrophoresis (DEP) has been widely used to manipulate, separate, and concentrate microscale particles. Unfortunately, DEP force is difficult to be used in regard to the manipulation of nanoscale molecules/particles. For manipulation of 50- to 100-nm particles, the electrical field strength must be higher than 3 × 106 V/m, and with a low applied voltage of 10 Vp-p, the electrode gap needs to be reduced to submicrons. Our research consists of a novel and simple approach, using a several tens micrometers scale electrode (low cost and easy to fabricate) to generate a dielectrophoretic microparticle assembly to form nanogaps with a locally amplified alternating current (AC) electric field gradient, which is used to rapidly trap nanocolloids. The results show that the amplified DEP force could effectively trap 20-nm colloids in the nanogaps between the 5-μm particle aggregates. The concentration factor at the local detection region was shown to be approximately 5 orders of magnitude higher than the bulk solution. This approach was also successfully used in bead-based surface-enhanced Raman spectroscopy (SERS) for the rapid identification of bacteria from diluted blood. PMID:25024685

  10. Exploring the binding of 4-thiothymidine with human serum albumin by spectroscopy, atomic force microscopy, and molecular modeling methods.

    PubMed

    Zhang, Juling; Gu, Huaimin; Zhang, Xiaohui

    2014-01-30

    The interaction of 4-thiothymidine (S(4)TdR) with human serum albumin (HSA) was studied by equilibrium dialysis under normal physiological conditions. In this work, the mechanism of the interaction between S(4)TdR and human serum albumin (HSA) was exploited by fluorescence, UV, CD circular, and SERS spectroscopic. Fluorescence and UV spectroscopy suggest that HSA intensities are significantly decreased when adding S(4)TdR to HAS, and the quenching mechanism of the fluorescence is static. Also, the ΔG, ΔH, and ΔS values across temperature indicated that hydrophobic interaction was the predominant binding force. The CD circular results show that there is little change in the secondary structure of HSA except the environment of amino acid changes when adding S(4)TdR to HSA. The surface-enhanced Raman scattering (SERS) shows that the interaction between S(4)TdR and HSA can be achieved through different binding sites which are probably located in the II A and III A hydrophobic pockets of HSA which correspond to Sudlow's I and II binding sites. In addition, the molecular modeling displays that S(4)TdR-HSA complex is stabilized by hydrophobic forces, which result from amino acid residues. The atomic force microscopy results revealed that the single HSA molecular dimensions were larger after interaction of 4-thiothymidine. This work would be useful to understand the state of the transportation, distribution, and metabolism of the anticancer drugs in the human body, and it could provide a useful biochemistry parameter for the development of new anti-cancer drugs and research of pharmacology mechanisms. PMID:24370945

  11. Influence of smectite suspension structure on sheet orientation in dry sediments: XRD and AFM applications.

    PubMed

    Zbik, Marek S; Frost, Ray L

    2010-06-15

    The structure-building phenomena within clay aggregates are governed by forces acting between clay particles. Measurements of such forces are important to understand in order to manipulate the aggregate structure for applications such as dewatering of mineral processing tailings. A parallel particle orientation is required when conducting XRD investigation on the oriented samples and conduct force measurements acting between basal planes of clay mineral platelets using atomic force microscopy (AFM). To investigate how smectite clay platelets were oriented on silicon wafer substrate when dried from suspension range of methods like SEM, XRD and AFM were employed. From these investigations, we conclude that high clay concentrations and larger particle diameters (up to 5 microm) in suspension result in random orientation of platelets in the substrate. The best possible laminar orientation in the clay dry film, represented in the XRD 001/020 intensity ratio of 47 was obtained by drying thin layers from 0.02 wt.% clay suspensions of the natural pH. Conducted AFM investigations show that smectite studied in water based electrolytes show very long-range repulsive forces lower in strength than electrostatic forces from double-layer repulsion. It was suggested that these forces may have structural nature. Smectite surface layers rehydrate in water environment forms surface gel with spongy and cellular texture which cushion approaching AFM probe. This structural effect can be measured in distances larger than 1000 nm from substrate surface and when probe penetrate this gel layer, structural linkages are forming between substrate and clay covered probe. These linkages prevent subsequently smooth detachments of AFM probe on way back when retrieval. This effect of tearing new formed structure apart involves larger adhesion-like forces measured in retrieval. It is also suggested that these effect may be enhanced by the nano-clay particles interaction.

  12. Thermal Infrared Spectroscopy of Atmospheric Species Critical to Radiative Forcing of Earth's Climate

    NASA Astrophysics Data System (ADS)

    Brasunas, J. C., Jr.; Kostiuk, T.; Livengood, T. A.; Hewagama, T.; Kolasinski, J. R.

    2014-12-01

    Thermal-infrared (from about 6 to 100 or more microns wavelength), emission-mode Fourier transform spectrometer (FTS) systems acquire radiometric spectra for diurnal diagnostics of atmospheric properties. We have been funded through NASA's Planetary Instrument Definition and Development Program (PIDDP) to develop CIRS-lite as a lightweight successor to the Goddard-developed Cassini CIRS FTS currently operating in Saturn orbit. CIRS-lite also has promise for Earth science due to its modest mass, power and volume requirements and novel technical capabilities. For Earth, CIRS-lite supports the characterization of climate radiative forcing, including trace species measurements such as methane. Detection capability beyond the typical limit of HgCdTe focal planes (about 16 microns wavelength) enables a more complete characterization of the greenhouse effect. As for trace-species quantification, a beyond-HgCdTe focal plane permits characterization of water without overlapping lines from other species, enabling better measurements of these other species such as methane at wavelengths reachable by HgCdTe.

  13. Communication: Does force spectroscopy of biomolecules probe their intrinsic dynamic properties?

    SciTech Connect

    Makarov, Dmitrii E.

    2014-12-28

    In single-molecule pulling experiments, the molecule of interest is attached to a much larger object such as an atomic force microscope tip or a micrometer sized bead. The measured dynamics of molecular transitions is therefore affected by the hydrodynamic drag on the pulling instrument itself. By considering the transitions within the combined system (the molecule and the instrument), it is shown here that two distinct physical regimes exist: when the intrinsic stiffness of the molecule is greater than that of the linker connecting the molecule to the pulling setup then the pulling experiment probes the intrinsic dynamics of the molecule with only relatively small (and quantifiable) corrections resulting from the pulling setup. In contrast, when the stiffness of the linker exceeds that of the molecule, the molecular transition in question involves concerted motion of the molecule and the pulling setup and the hydrodynamic drag on the pulling instrument becomes the dominant source of friction along the molecular reaction coordinate. An analytical formula interpolating between these two cases is further derived. These results explain recent conflicting observations where some single-molecule pulling measurements report anomalously low diffusion coefficients along molecular reaction coordinates while others do not.

  14. Application of atomic force microscopy for characteristics of single intermolecular interactions.

    PubMed

    Safenkova, I V; Zherdev, A V; Dzantievf, B B

    2012-12-01

    Atomic force microscopy (AFM) can be used to make measurements in vacuum, air, and water. The method is able to gather information about intermolecular interaction forces at the level of single molecules. This review encompasses experimental and theoretical data on the characterization of ligand-receptor interactions by AFM. The advantage of AFM in comparison with other methods developed for the characterization of single molecular interactions is its ability to estimate not only rupture forces, but also thermodynamic and kinetic parameters of the rupture of a complex. The specific features of force spectroscopy applied to ligand-receptor interactions are examined in this review from the stage of the modification of the substrate and the cantilever up to the processing and interpretation of the data. We show the specificities of the statistical analysis of the array of data based on the results of AFM measurements, and we discuss transformation of data into thermodynamic and kinetic parameters (kinetic dissociation constant, Gibbs free energy, enthalpy, and entropy). Particular attention is paid to the study of polyvalent interactions, where the definition of the constants is hampered due to the complex stoichiometry of the reactions.

  15. Electrical characterization of grain boundaries of CZTS thin films using conductive atomic force microscopy techniques

    SciTech Connect

    Muhunthan, N.; Singh, Om Pal; Toutam, Vijaykumar; Singh, V.N.

    2015-10-15

    Graphical abstract: Experimental setup for conducting AFM (C-AFM). - Highlights: • Cu{sub 2}ZnSnS{sub 4} (CZTS) thin film was grown by reactive co-sputtering. • The electronic properties were probed using conducting atomic force microscope, scanning Kelvin probe microscopy and scanning capacitance microscopy. • C-AFM current flow mainly through grain boundaries rather than grain interiors. • SKPM indicated higher potential along the GBs compared to grain interiors. • The SCM explains that charge separation takes place at the interface of grain and grain boundary. - Abstract: Electrical characterization of grain boundaries (GB) of Cu-deficient CZTS (Copper Zinc Tin Sulfide) thin films was done using atomic force microscopic (AFM) techniques like Conductive atomic force microscopy (CAFM), Kelvin probe force microscopy (KPFM) and scanning capacitance microscopy (SCM). Absorbance spectroscopy was done for optical band gap calculations and Raman, XRD and EDS for structural and compositional characterization. Hall measurements were done for estimation of carrier mobility. CAFM and KPFM measurements showed that the currents flow mainly through grain boundaries (GB) rather than grain interiors. SCM results showed that charge separation mainly occurs at the interface of grain and grain boundaries and not all along the grain boundaries.

  16. Coffee Cup Atomic Force Microscopy

    ERIC Educational Resources Information Center

    Ashkenaz, David E.; Hall, W. Paige; Haynes, Christy L.; Hicks, Erin M.; McFarland, Adam D.; Sherry, Leif J.; Stuart, Douglas A.; Wheeler, Korin E.; Yonzon, Chanda R.; Zhao, Jing; Godwin, Hilary A.; Van Duyne, Richard P.

    2010-01-01

    In this activity, students use a model created from a coffee cup or cardstock cutout to explore the working principle of an atomic force microscope (AFM). Students manipulate a model of an AFM, using it to examine various objects to retrieve topographic data and then graph and interpret results. The students observe that movement of the AFM…

  17. Application of catalyst-free click reactions in attaching affinity molecules to tips of atomic force microscopy for detection of protein biomarkers.

    PubMed

    Senapati, Subhadip; Manna, Saikat; Lindsay, Stuart; Zhang, Peiming

    2013-11-26

    Atomic force microscopy (AFM) has been extensively used in studies of biological interactions. Particularly, AFM based force spectroscopy and recognition imaging can sense biomolecules on a single molecule level, having great potential to become a tool for molecular diagnostics in clinics. These techniques, however, require affinity molecules to be attached to AFM tips in order to specifically detect their targets. The attachment chemistry currently used on silicon tips involves multiple steps of reactions and moisture sensitive chemicals, such as (3-aminopropyl)triethoxysilane (APTES) and N-hydroxysuccinimide (NHS) ester, making the process difficult to operate in aqueous solutions. In the present study, we have developed a user-friendly protocol to functionalize the AFM tips with affinity molecules. A key feature of it is that all reactions are carried out in aqueous solutions. In summary, we first synthesized a molecular anchor composed of cyclooctyne and silatrane for introduction of a chemically reactive function to AFM tips and a bifunctional polyethylene glycol linker that harnesses two orthogonal click reactions, copper free alkyne-azide cycloaddition and thiol-vinylsulfone Michael addition, for attaching affinity molecules to AFM tips. The attachment chemistry was then validated by attaching antithrombin DNA aptamers and cyclo-RGD peptides to silicon nitride (SiN) tips, respectively, and measuring forces of unbinding these affinity molecules from their protein cognates human α-thrombin and human α5β1-integrin immobilized on mica surfaces. In turn, we used the same attachment chemistry to functionalize silicon tips with the same affinity molecules for AFM based recognition imaging, showing that the disease-relevant biomarkers such as α-thrombin and α5β1-integrin can be detected with high sensitivity and specificity by the single molecule technique. These studies demonstrate the feasibility of our attachment chemistry for the use in functionalization

  18. Application of Catalyst-free Click Reactions in Attaching Affinity Molecules to Tips of Atomic Force Microscopy for Detection of Protein Biomarkers

    PubMed Central

    Senapati, Subhadip; Manna, Saikat; Lindsay, Stuart; Zhang, Peiming

    2013-01-01

    Atomic Force Microscopy (AFM) has been extensively used in studies of biological interactions. Particularly, AFM based force spectroscopy and recognition imaging can sense biomolecules on a single molecule level, having great potential to become a tool for molecular diagnostics in clinics. These techniques, however, require affinity molecules to be attached to AFM tips in order to specifically detect their targets. The attachment chemistry currently used on silicon tips involves multiple steps of reactions and moisture sensitive chemicals, such as (3-aminopropyl)triethoxysilane (APTES) and N-hydroxysuccinimide (NHS) ester, making the process difficult to operate in aqueous solutions. In the present study, we have developed a user-friendly protocol to functionalize the AFM tips with affinity molecules. A key feature of it is that all reactions are carried out in aqueous solutions. In summary, we first synthesized a molecular anchor composed of cyclooctyne and silatrane for introduction of a chemically reactive function to AFM tips and a bi-functional polyethylene glycol linker that harnesses two orthogonal click reactions, copper free alkyne-azide cycloaddition and thiol-vinylsulfone Michael addition, for attaching affinity molecules to AFM tips. The attachment chemistry was then validated by attaching anti-thrombin DNA aptamers and cyclo-RGD peptides to silicon nitride (SiN) tips respectively, and measuring forces of unbinding these affinity molecules from their protein cognates human α-thrombin and human α5β1-integrin immobilized on mica surfaces. In turn, we used the same attachment chemistry to functionalize silicon tips with the same affinity molecules for AFM based recognition imaging, showing that the disease-relevant biomarkers such as α-thrombin and α5β1-integrin can be detected with high sensitivity and specificity by the single molecule technique. These studies demonstrate the feasibility of our attachment chemistry for the use in functionalization

  19. Hydration states of AFm cement phases

    SciTech Connect

    Baquerizo, Luis G.; Matschei, Thomas; Scrivener, Karen L.; Saeidpour, Mahsa; Wadsö, Lars

    2015-07-15

    The AFm phase, one of the main products formed during the hydration of Portland and calcium aluminate cement based systems, belongs to the layered double hydrate (LDH) family having positively charged layers and water plus charge-balancing anions in the interlayer. It is known that these phases present different hydration states (i.e. varying water content) depending on the relative humidity (RH), temperature and anion type, which might be linked to volume changes (swelling and shrinkage). Unfortunately the stability conditions of these phases are insufficiently reported. This paper presents novel experimental results on the different hydration states of the most important AFm phases: monocarboaluminate, hemicarboaluminate, strätlingite, hydroxy-AFm and monosulfoaluminate, and the thermodynamic properties associated with changes in their water content during absorption/desorption. This data opens the possibility to model the response of cementitious systems during drying and wetting and to engineer systems more resistant to harsh external conditions.

  20. AFM-assisted fabrication of thiol SAM pattern with alternating quantified surface potential

    PubMed Central

    2011-01-01

    Thiol self-assembled monolayers (SAMs) are widely used in many nano- and bio-technology applications. We report a new approach to create and characterize a thiol SAMs micropattern with alternating charges on a flat gold-coated substrate using atomic force microscopy (AFM) and Kelvin probe force microscopy (KPFM). We produced SAMs-patterns made of alternating positively charged, negatively charged, and hydrophobic-terminated thiols by an automated AFM-assisted manipulation, or nanografting. We show that these thiol patterns possess only small topographical differences as revealed by AFM, and distinguished differences in surface potential (20-50 mV), revealed by KPFM. The pattern can be helpful in the development of biosensor technologies, specifically for selective binding of biomolecules based on charge and hydrophobicity, and serve as a model for creating surfaces with quantified alternating surface potential distribution. PMID:21711703

  1. Atomic Force Microscope

    SciTech Connect

    Day, R.D.; Russell, P.E.

    1988-12-01

    The Atomic Force Microscope (AFM) is a recently developed instrument that has achieved atomic resolution imaging of both conducting and non- conducting surfaces. Because the AFM is in the early stages of development, and because of the difficulty of building the instrument, it is currently in use in fewer than ten laboratories worldwide. It promises to be a valuable tool for obtaining information about engineering surfaces and aiding the .study of precision fabrication processes. This paper gives an overview of AFM technology and presents plans to build an instrument designed to look at engineering surfaces.

  2. High throughput nanofabrication of silicon nanowire and carbon nanotube tips on AFM probes by stencil-deposited catalysts.

    PubMed

    Engstrom, Daniel S; Savu, Veronica; Zhu, Xueni; Bu, Ian Y Y; Milne, William I; Brugger, Juergen; Boggild, Peter

    2011-04-13

    A new and versatile technique for the wafer scale nanofabrication of silicon nanowire (SiNW) and multiwalled carbon nanotube (MWNT) tips on atomic force microscope (AFM) probes is presented. Catalyst material for the SiNW and MWNT growth was deposited on prefabricated AFM probes using aligned wafer scale nanostencil lithography. Individual vertical SiNWs were grown epitaxially by a catalytic vapor-liquid-solid (VLS) process and MWNTs were grown by a plasma-enhanced chemical vapor (PECVD) process on the AFM probes. The AFM probes were tested for imaging micrometers-deep trenches, where they demonstrated a significantly better performance than commercial high aspect ratio tips. Our method demonstrates a reliable and cost-efficient route toward wafer scale manufacturing of SiNW and MWNT AFM probes. PMID:21446752

  3. Characterization of single 1.8-nm Au nanoparticle attachments on AFM tips for single sub-4-nm object pickup

    NASA Astrophysics Data System (ADS)

    Cheng, Hui-Wen; Chang, Yuan-Chih; Tang, Song-Nien; Yuan, Chi-Tsu; Tang, Jau; Tseng, Fan-Gang

    2013-11-01

    This paper presents a novel method for the attachment of a 1.8-nm Au nanoparticle (Au-NP) to the tip of an atomic force microscopy (AFM) probe through the application of a current-limited bias voltage. The resulting probe is capable of picking up individual objects at the sub-4-nm scale. We also discuss the mechanisms involved in the attachment of the Au-NP to the very apex of an AFM probe tip. The Au-NP-modified AFM tips were used to pick up individual 4-nm quantum dots (QDs) using a chemically functionalized method. Single QD blinking was reduced considerably on the Au-NP-modified AFM tip. The resulting AFM tips present an excellent platform for the manipulation of single protein molecules in the study of single protein-protein interactions.

  4. Fabrication of carbon nanotube AFM probes using the Langmuir-Blodgett technique.

    PubMed

    Lee, Jae-Hyeok; Kang, Won-Seok; Choi, Bung-Sam; Choi, Sung-Wook; Kim, Jae-Ho

    2008-09-01

    Carbon nanotube (CNT)-tipped atomic force microscopy (AFM) probes have shown a significant potential for obtaining high-resolution imaging of nanostructure and biological materials. In this paper, we report a simple method to fabricate single-walled carbon nanotube (SWNT) nanoprobes for AFM using the Langmuir-Blodgett (LB) technique. Thiophenyl-modified SWNTs (SWNT-SHs) through amidation of SWNTs in chloroform allowed to be spread and form a stable Langmuir monolayer at the water/air interface. A simple two-step transfer process was used: (1) dipping conventional AFM probes into the Langmuir monolayer and (2) lifting the probes from the water surface. This results in the attachment of SWNTs onto the tips of AFM nanoprobes. We found that the SWNTs assembled on the nanoprobes were well-oriented and robust enough to maintain their shape and direction even after successive scans. AFM measurements of a nano-porous alumina substrate and deoxyribonucleic acid using SWNT-modified nanoprobes revealed that the curvature diameter of the nanoprobes was less than 3 nm and a fine resolution was obtained than that from conventional AFM probes. We also demonstrate that the LB method is a scalable process capable of simultaneously fabricating a large number of SWNT-modified nanoprobes.

  5. Probing molecular interaction in ionic liquids by low frequency spectroscopy: Coulomb energy, hydrogen bonding and dispersion forces.

    PubMed

    Fumino, Koichi; Reimann, Sebastian; Ludwig, Ralf

    2014-10-28

    Ionic liquids are defined as salts composed solely of ions with melting points below 100 °C. These remarkable liquids have unique and fascinating properties and offer new opportunities for science and technology. New combinations of ions provide changing physical properties and thus novel potential applications for this class of liquid materials. To a large extent, the structure and properties of ionic liquids are determined by the intermolecular interaction between anions and cations. In this perspective we show that far infrared and terahertz spectroscopy are suitable methods for studying the cation-anion interaction in these Coulomb fluids. The interpretation of the measured low frequency spectra is supported by density functional theory calculations and molecular dynamics simulations. We present results for selected aprotic and protic ionic liquids and their mixtures with molecular solvents. In particular, we focus on the strength and type of intermolecular interaction and how both parameters are influenced by the character of the ions and their combinations. We show that the total interaction between cations and anions is a result of a subtle balance between Coulomb forces, hydrogen bonds and dispersion forces. For protic ionic liquids we could measure distinct vibrational modes in the low frequency spectra indicating clearly the cation-anion interaction characterized by linear and medium to strong hydrogen bonds. Using isotopic substitution we have been able to dissect frequency shifts related to pure interaction strength between cations and anions and to different reduced masses only. In this context we also show how these different types of interaction may influence the physical properties of ionic liquids such as the melting point, viscosity or enthalpy of vaporization. Furthermore we demonstrate that low frequency spectroscopy can also be used for studying ion speciation. Low vibrational features can be assigned to contact ion pairs and solvent separated

  6. Visualization of internal structure of banana starch granule through AFM.

    PubMed

    Peroni-Okita, Fernanda H G; Gunning, A Patrick; Kirby, Andrew; Simão, Renata A; Soares, Claudinéia A; Cordenunsi, Beatriz R

    2015-09-01

    Atomic force microscopy (AFM) is a high resolution technique for studying the external and internal structures of starch granules. For this purpose granules were isolated from bananas and embedded in a non-penetrating resin. To achieve image contrast of the ultrastructure, the face of the cut blocks were wetted in steam and force modulation mode imaging was used. Images of starch from green bananas showed large variation of height across the granule due to a locational specific absorption of water and swelling of amorphous regions; the data reveal that the center of the granules are structurally different and have different viscoelastic properties. Images of starches from ripe bananas showed an even greater different level of organization: absence of growth rings around the hilum; the central region of the granule is richer in amylose; very porous surface with round shaped dark structures; the size of blocklets are larger than the green fruits. PMID:26005137

  7. Mechanical properties of poly(dimethylsiloxane)-block-poly(2-methyloxazoline) polymersomes probed by atomic force microscopy.

    PubMed

    Jaskiewicz, Karmena; Makowski, Marcin; Kappl, Michael; Landfester, Katharina; Kroeger, Anja

    2012-08-28

    Poly(dimethylsiloxane)-block-poly(2-methyloxazoline) (PDMS-b-PMOXA) vesicles were characterized by a combination of dynamic light scattering (DLS), cryogenic transmission electron microscopy (cryo-TEM), and atomic force microscopy imaging and force spectroscopy (AFM). From DLS data, a hydrodynamic radius of ~150 nm was determined, and cryo-TEM micrographs revealed a bilayer thickness of ~16 nm. In AFM experiments on a silicon wafer substrate, adsorption led to a stable spherical caplike conformation of the polymersomes, whereas on mica, adsorption resulted also in vesicle fusion and formation of bilayer patches or multilayer stacks. This indicates a delicate balance between the mechanical stability of PDMS-b-PMOXA polymersomes on one hand and the driving forces for spreading on the other. A Young's modulus of 17 ± 11 MPa and a bending modulus of 7 ± 5 × 10(-18) J were derived from AFM force spectroscopy measurements. Therefore, the elastic response of the PDMS-b-PMOXA polymersomes to external stimuli is much closer to that of lipid vesicles compared to other types of polymersomes, such as polystyrene-block-poly(acrylic acid) (PS-b-PAA).

  8. AFM probes fabricated with masked maskless combined anisotropic etching and p+ surface doping

    NASA Astrophysics Data System (ADS)

    Han, Jianqiang; Li, Xinxin; Bao, Haifei; Zuo, Guomin; Wang, Yuelin; Feng, Fei; Yu, Zhenyin; Ge, Xiaohong

    2006-02-01

    The paper presents a newly developed high-yield micro-fabrication technology for single-crystalline silicon atomic force microscope (AFM) probes. Both the tips and the cantilevers are simultaneously formed by a masked-maskless combined anisotropic etching process. Compared to a conventional tip-to-cantilever sequential fabrication scheme, this tip-and-cantilever simultaneous formation can effectively increase fabrication yield by avoiding the tips damaged during the following processed photolithographic steps for defining the cantilevers. By heavy boron doping at the surface, the conductive AFM probe provides an electrical path to the electric ground of the AFM that helps to eliminate the electrostatic accumulation of charges and, therefore, eliminate undesirable electrostatic forces between the probes and the samples. A fabrication yield as high as 90% has been obtained for the AFM probes for 4 inch wafers. The tips after oxidation-sharpening treatment generally have a radius of 10-30 nm. The cantilever spring constant can be well controlled in the range of 0.025-40 N m-1. High-quality sample scanning results with the formed AFM probes are obtained with a slightly better resolution than that from commercial probes without surface conductive treatment.

  9. Effect of AFM probe geometry on visco-hyperelastic characterization of soft materials

    NASA Astrophysics Data System (ADS)

    Boccaccio, Antonio; Lamberti, Luciano; Papi, Massimiliano; De Spirito, Marco; Pappalettere, Carmine

    2015-08-01

    Atomic force microscopy (AFM) nanoindentation is very suited for nano- and microscale mechanical characterization of soft materials. Although the structural response of polymeric networks that form soft matter depends on viscous effects caused by the relative slippage of polymeric chains, the usual assumption made in the AFM-based characterization is that the specimen behaves as a purely elastic material and viscous forces are negligible. However, for each geometric configuration of the AFM tip, there will be a limit indentation rate above which viscous effects must be taken into account to correctly determine mechanical properties. A parametric finite element study conducted on 12 geometric configurations of a blunt cone AFM tip (overall, the study included about 200 finite element analyses) allowed us to determine the limit indentation rate for each configuration. The selected tip dimensions cover commercially available products and account for changes in tip geometry caused by serial measurements. Nanoindentation rates cover typical experimental conditions set in AFM bio-measurements on soft matter. Viscous effects appear to be more significant in the case of sharper tips. This implies that, if quantitative data on sample viscosity are not available, using a rounded indenter and carrying out experiments below the limit indentation rate will allow errors in the determination of mechanical properties to be minimized.

  10. TAT peptide immobilization on gold surfaces: a comparison study with a thiolated peptide and alkylthiols using AFM, XPS, and FT-IRRAS.

    PubMed

    Cho, Youngnam; Ivanisevic, Albena

    2005-04-01

    A TAT peptide was used to functionalize a gold surface by three different methods: adsorption from solution, microcontact printing, and dip-pen nanolithography (DPN). The composition and structure of the modified gold was characterized by atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and Fourier transform -infrared reflection absorption spectroscopy (FT-IRRAS). We used two well-studied alkylthiols, mercaptohexadecanoic acid and 1-octadecanethiol, as a comparison in order to understand the structure of the TAT peptide monolayers prepared by the three methods. AFM studies allowed us to assess the homogeneity after each modification protocol. XPS was used to characterize the chemical composition of the gold surface after each functionalization procedure. The XPS results showed that surfaces modified with the TAT peptide by the three methods exhibit similar surface chemistry. Finally, FT-IRRAS experiments allowed us to conclude that the structure of the alkyl chains of the TAT peptides is fairly disordered and different after each procedure. Regardless of the type of surface functionalization method used, the monolayer of TAT peptide formed on the surface was of "liquidlike" nature.

  11. Local mobility in lipid domains of supported bilayers characterized by atomic force microscopy and fluorescence correlation spectroscopy.

    SciTech Connect

    Frankel, Daniel J.; Buranda, T.; Burns, Alan Richard

    2005-01-01

    Fluorescence correlation spectroscopy (FCS) is used to examine mobility of labeled probes at specific sites in supported bilayers consisting of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) lipid domains in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). Those sites are mapped beforehand with simultaneous atomic force microscopy and submicron confocal fluorescence imaging, allowing characterization of probe partitioning between gel DPPC and disordered liquid DOPC domains with corresponding topography of domain structure. We thus examine the relative partitioning and mobility in gel and disordered liquid phases for headgroup- and tailgroup-labeled GM1 ganglioside probes and for headgroup- and tailgroup-labeled phospholipid probes. For the GM1 probes, large differences in mobility between fluid and gel domains are observed; whereas unexpected mobility is observed in submicron gel domains for the phospholipid probes. We attribute the latter to domain heterogeneities that could be induced by the probe. Furthermore, fits to the FCS data for the phospholipid probes in the DOPC fluid phase require two components (fast and slow). Although proximity to the glass substrate may be a factor, local distortion of the probe by the fluorophore could also be important. Overall, we observe nonideal aspects of phospholipid probe mobility and partitioning that may not be restricted to supported bilayers.

  12. MDM2–MDM4 molecular interaction investigated by atomic force spectroscopy and surface plasmon resonance

    PubMed Central

    Moscetti, Ilaria; Teveroni, Emanuela; Moretti, Fabiola; Bizzarri, Anna Rita; Cannistraro, Salvatore

    2016-01-01

    Murine double minute 2 (MDM2) and 4 (MDM4) are known as the main negative regulators of p53, a tumor suppressor. They are able to form heterodimers that are much more effective in the downregulation of p53. Therefore, the MDM2–MDM4 complex could be a target for promising therapeutic restoration of p53 function. To this aim, a deeper understanding of the molecular mechanisms underlining the heterodimerization is needed. The kinetic and thermodynamic characterization of the MDM2–MDM4 complex was performed with two complementary approaches: atomic force spectroscopy and surface plasmon resonance. Both techniques revealed an equilibrium dissociation constant (KD) in the micromolar range for the MDM2–MDM4 heterodimer, similar to related complexes involved in the p53 network. Furthermore, the MDM2–MDM4 complex is characterized by a relatively high free energy, through a single energy barrier, and by a lifetime in the order of tens of seconds. New insights into the MDM2–MDM4 interaction could be highly important for developing innovative anticancer drugs focused on p53 reactivation. PMID:27621617

  13. MDM2–MDM4 molecular interaction investigated by atomic force spectroscopy and surface plasmon resonance

    PubMed Central

    Moscetti, Ilaria; Teveroni, Emanuela; Moretti, Fabiola; Bizzarri, Anna Rita; Cannistraro, Salvatore

    2016-01-01

    Murine double minute 2 (MDM2) and 4 (MDM4) are known as the main negative regulators of p53, a tumor suppressor. They are able to form heterodimers that are much more effective in the downregulation of p53. Therefore, the MDM2–MDM4 complex could be a target for promising therapeutic restoration of p53 function. To this aim, a deeper understanding of the molecular mechanisms underlining the heterodimerization is needed. The kinetic and thermodynamic characterization of the MDM2–MDM4 complex was performed with two complementary approaches: atomic force spectroscopy and surface plasmon resonance. Both techniques revealed an equilibrium dissociation constant (KD) in the micromolar range for the MDM2–MDM4 heterodimer, similar to related complexes involved in the p53 network. Furthermore, the MDM2–MDM4 complex is characterized by a relatively high free energy, through a single energy barrier, and by a lifetime in the order of tens of seconds. New insights into the MDM2–MDM4 interaction could be highly important for developing innovative anticancer drugs focused on p53 reactivation.

  14. Analysis of Snow Albedo, Grain Size and Radiative Forcing based on the Airborne Snow Observatory (ASO) Imaging Spectroscopy Data

    NASA Astrophysics Data System (ADS)

    Seidel, F. C.; Painter, T. H.

    2013-12-01

    Climate is expected to be most vulnerable in mountainous and arctic regions where the atmosphere and the hydrosphere are directly linked to the cryosphere. A combination of modeling and large-scale observational efforts is required to investigate related scientific questions. NASA's Airborne Snow Observatory (ASO) at the Jet Propulsion Laboratory addresses some of these needs by establishing new quantitative observational capabilities in regional mapping of mountain snow properties. In addition, ASO's key products showed that we are able to achieve societal benefits by improving water resources management. We will show the first analysis of snow optical products (albedo, grain size, and radiative forcing) from the spring 2013 ASO campaign in the Sierra Nevada, CA, USA. In addition, we will present the retrieval methods used to derive these products based on airborne imaging spectroscopy, LiDAR, as well as radiative transfer models. The preliminary findings provide new important insights into the temporal and spatial aspects of Western US mountain snow and its melt.

  15. The structure of high-methoxyl sugar acid gels of citrus pectin as determined by AFM

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Images of native high methoxyl sugar acid gels (HMSAG) were obtained by atomic force microscopy (AFM) in the Tapping ModeTM. Electronic thinning of the pectin strands to one pixel wide allowed the pectin network to be viewed in the absence of variable strand widths related to preferentially solvate...

  16. Accurate calibration and uncertainty estimation of the normal spring constant of various AFM cantilevers.

    PubMed

    Song, Yunpeng; Wu, Sen; Xu, Linyan; Fu, Xing

    2015-03-10

    Measurement of force on a micro- or nano-Newton scale is important when exploring the mechanical properties of materials in the biophysics and nanomechanical fields. The atomic force microscope (AFM) is widely used in microforce measurement. The cantilever probe works as an AFM force sensor, and the spring constant of the cantilever is of great significance to the accuracy of the measurement results. This paper presents a normal spring constant calibration method with the combined use of an electromagnetic balance and a homemade AFM head. When the cantilever presses the balance, its deflection is detected through an optical lever integrated in the AFM head. Meanwhile, the corresponding bending force is recorded by the balance. Then the spring constant can be simply calculated using Hooke's law. During the calibration, a feedback loop is applied to control the deflection of the cantilever. Errors that may affect the stability of the cantilever could be compensated rapidly. Five types of commercial cantilevers with different shapes, stiffness, and operating modes were chosen to evaluate the performance of our system. Based on the uncertainty analysis, the expanded relative standard uncertainties of the normal spring constant of most measured cantilevers are believed to be better than 2%.

  17. AFM and EDX Study of Self Assembled Pt Nanostructures on PEDOT Thin Films under Ambient Conditions

    NASA Astrophysics Data System (ADS)

    Senevirathne, Indrajith; Mohney, Austin; Buchheit, Joshua; Goonewardene, Anura

    2011-03-01

    Noble metal nanostructure systems on conductive polymer thin films under ambient conditions are interesting due to their use in BioMEMS and hybrid systems further and considering the physics of the polymer - metal interactions The observed nanostructures have deformed spherical shape. The Pt was magnetron sputter deposited at RT (300K), PEDOT Baytron P 60nm thick, spin coated on glass slides cleaned with acetone and IPA. The system was studied using ambient IC mode Atomic Force Microscopy (AFM) for its structure. Elemental composition/distribution of the system was measured with Energy Dispersive X ray Spectroscopy (EDX). Pt nanostructures on the surface observed to be likely Volmer - Weber growth mode At Pt coverage of 120 ML, nanostructures had a mean diameter of 32 nm and mean height of 5 nm. When annealing at 15min at 473K systems changes to smaller nanostructures coexisting with bigger structures of mean diameter of 120 nm and mean height of 36 nm. Elemental/morphological variations when annealed at successively higher temperatures were also investigated. NSF Grant #: 0923047 and PASSHE FPDC (LOU # 2010-LHU-03).

  18. Two-Dimensional Measurement of n+-p Asymmetrical Junctions in Multicrystalline Silicon Solar Cells Using AFM-Based Electrical Techniques with Nanometer Resolution: Preprint

    SciTech Connect

    Jiang, C. S.; Moutinho, H. R.; Li, J. V.; Al-Jassim, M. M.; Heath, J. T.

    2011-07-01

    Lateral inhomogeneities of modern solar cells demand direct electrical imaging with nanometer resolution. We show that atomic force microscopy (AFM)-based electrical techniques provide unique junction characterizations, giving a two-dimensional determination of junction locations. Two AFM-based techniques, scanning capacitance microscopy/spectroscopy (SCM/SCS) and scanning Kelvin probe force microscopy (SKPFM), were significantly improved and applied to the junction characterizations of multicrystalline silicon (mc-Si) cells. The SCS spectra were taken pixel by pixel by precisely controlling the tip positions in the junction area. The spectra reveal distinctive features that depend closely on the position relative to the electrical junction, which allows us to indentify the electrical junction location. In addition, SKPFM directly probes the built-in potential over the junction area modified by the surface band bending, which allows us to deduce the metallurgical junction location by identifying a peak of the electric field. Our results demonstrate resolutions of 10-40 nm, depending on the techniques (SCS or SKPFM). These direct electrical measurements with nanometer resolution and intrinsic two-dimensional capability are well suited for investigating the junction distribution of solar cells with lateral inhomogeneities.

  19. Image Analysis and Length Estimation of Biomolecules Using AFM

    PubMed Central

    Sundstrom, Andrew; Cirrone, Silvio; Paxia, Salvatore; Hsueh, Carlin; Kjolby, Rachel; Gimzewski, James K.; Reed, Jason; Mishra, Bud

    2014-01-01

    There are many examples of problems in pattern analysis for which it is often possible to obtain systematic characterizations, if in addition a small number of useful features or parameters of the image are known a priori or can be estimated reasonably well. Often, the relevant features of a particular pattern analysis problem are easy to enumerate, as when statistical structures of the patterns are well understood from the knowledge of the domain. We study a problem from molecular image analysis, where such a domain-dependent understanding may be lacking to some degree and the features must be inferred via machine-learning techniques. In this paper, we propose a rigorous, fully automated technique for this problem. We are motivated by an application of atomic force microscopy (AFM) image processing needed to solve a central problem in molecular biology, aimed at obtaining the complete transcription profile of a single cell, a snapshot that shows which genes are being expressed and to what degree. Reed et al. (“Single molecule transcription profiling with AFM,” Nanotechnology, vol. 18, no. 4, 2007) showed that the transcription profiling problem reduces to making high-precision measurements of biomolecule backbone lengths, correct to within 20–25 bp (6–7.5 nm). Here, we present an image processing and length estimation pipeline using AFM that comes close to achieving these measurement tolerances. In particular, we develop a biased length estimator on trained coefficients of a simple linear regression model, biweighted by a Beaton–Tukey function, whose feature universe is constrained by James–Stein shrinkage to avoid overfitting. In terms of extensibility and addressing the model selection problem, this formulation subsumes the models we studied. PMID:22759526

  20. Simulation of CNT-AFM tip based on finite element analysis for targeted probe of the biological cell

    NASA Astrophysics Data System (ADS)

    Yousefi, Amin Termeh; Mahmood, Mohamad Rusop; Miyake, Mikio; Ikeda, Shoichiro

    2016-07-01

    Carbon nanotubes (CNTs) are potentially ideal tips for atomic force microscopy (AFM) due to the robust mechanical properties, nano scale diameter and also their ability to be functionalized by chemical and biological components at the tip ends. This contribution develops the idea of using CNTs as an AFM tip in computational analysis of the biological cell's. Finite element analysis employed for each section and displacement of the nodes located in the contact area was monitored by using an output database (ODB). This reliable integration of CNT-AFM tip process provides a new class of high performance nanoprobes for single biological cell analysis.

  1. Applications of atomic force microscopy to the studies of biomaterials in biomolecular systems

    NASA Astrophysics Data System (ADS)

    Ma, Xiang

    Atomic force microscopy (AFM) is a unique tool for the studies of nanoscale structures and interactions. In this dissertation, I applied AFM to study transitions among multiple states of biomaterials in three different microscopic biomolecular systems: MukB-dependent DNA condensation, holdfast adhesion, and virus elasticity. To elucidate the mechanism of MukB-dependent DNA condensation, I have studied the conformational changes of MukB proteins as indicators for the strength of interactions between MukB, DNA and other molecular factors, such as magnesium and ParC proteins, using high-resolution AFM imaging. To determine the physical origins of holdfast adhesion, I have investigated the dynamics of adhesive force development of the holdfast, employing AFM force spectroscopy. By measuring rupture forces between the holdfast and the substrate, I showed that the holdfast adhesion is strongly time-dependent and involves transformations at multiple time scales. Understanding the mechanisms of adhesion force development of the holdfast will be critical for future engineering of holdfasts properties for various applications. Finally, I have examined the elasticity of self-assembled hepatitis B virus-like particles (HBV VLPs) and brome mosaic virus (BMV) in response to changes of pH and salinity, using AFM nanoindentation. The distributions of elasticity were mapped on a single particle level and compared between empty, RNA- and gold-filled HBV VLPs. I found that a single HBV VLP showed heterogeneous distribution of elasticity and a two-step buckling transition, suggesting a discrete property of HBV capsids. For BMV, I have showed that viruses containing different RNA molecules can be distinguished by mechanical measurements, while they are indistinguishable by morphology. I also studied the effect of pH on the elastic behaviors of three-particle BMV and R3/4 BMV. This study can yield insights into RNA presentation/release mechanisms, and could help us to design novel drug

  2. Atomic force microscopy imaging using a tip-on-chip: Opening the door to integrated near field nanotools

    SciTech Connect

    Hayton, J.; Polesel-Maris, J.; Thoyer, F.; Lubin, C.; Cousty, J.; Demadrille, R.; Grevin, B.; Brun, M.

    2010-09-15

    We describe in detail how atomic force microscopy (AFM) images can be routinely achieved with macroscopic silicon-based chips integrating mesoscopic tips, paving the way for the development of new near field devices combining AFM imaging with any kind of functionality integrated on a chip. The chips have been glued at the end of the free prong of 100 kHz quartz tuning forks mounted in Qplus configuration. Numerical simulations by modal analysis have been carried out to clarify the nature of the vibration modes observed in the experimental spectra. It is shown that two low frequency modes can be used to drive the system and scan the surface with a great stability in amplitude modulation as well as in frequency modulation AFM under ultrahigh vacuum. The AFM capabilities are demonstrated through a series of examples including phase and dissipation contrast imaging, force spectroscopy measurements, and investigations of soft samples in weak interaction with the substrate. The lateral resolution with the tips grown by focused ion beam deposition already matches the one achieved in standard amplitude modulation mode AFM experiments.

  3. Adhesive force measurement between HOPG and zinc oxide as an indicator for interfacial bonding of carbon fiber composites.

    PubMed

    Patterson, Brendan A; Galan, Ulises; Sodano, Henry A

    2015-07-22

    Vertically aligned zinc oxide (ZnO) nanowires have recently been utilized as an interphase to increase the interfacial strength of carbon fiber composites. It was shown that the interaction between the carbon fiber and the ZnO nanowires was a critical parameter in adhesion; however, fiber based testing techniques are dominated by local defects and cannot be used to effectively study the bonding interaction directly. Here, the strength of the interface between ZnO and graphitic carbon is directly measured with atomic force microscopy (AFM) using oxygen plasma treated highly oriented pyrolytic graphite (HOPG) and an AFM tip coated with ZnO nanoparticles. X-ray photoelectron spectroscopy analysis is used to compare the surface chemistry of HOPG and carbon fiber and to quantify the presence of various oxygen functional groups. An indirect measurement of the interfacial strength is then performed through single fiber fragmentation testing (SFF) on functionalized carbon fibers coated with ZnO nanowires to validate the AFM measurements. The SFF and AFM methods showed the same correlation, demonstrating the capacity of the AFM method to study the interfacial properties in composite materials. Additionally, the chemical interactions between oxygen functional groups and the ionic structure of ZnO suggest that intermolecular forces at the interface are responsible for the strong interface.

  4. Direct visualization of the trimeric structure of the ASIC1a channel, using AFM imaging

    SciTech Connect

    Carnally, Stewart M.; Dev, Harveer S.; Stewart, Andrew P.; Barrera, Nelson P.; Van Bemmelen, Miguel X.; Schild, Laurent; Henderson, Robert M.; Edwardson, J.Michael

    2008-08-08

    There has been confusion about the subunit stoichiometry of the degenerin family of ion channels. Recently, a crystal structure of acid-sensing ion channel (ASIC) 1a revealed that it assembles as a trimer. Here, we used atomic force microscopy (AFM) to image unprocessed ASIC1a bound to mica. We detected a mixture of subunit monomers, dimers and trimers. In some cases, triple-subunit clusters were clearly visible, confirming the trimeric structure of the channel, and indicating that the trimer sometimes disaggregated after adhesion to the mica surface. This AFM-based technique will now enable us to determine the subunit arrangement within heteromeric ASICs.

  5. AFM and XPA data on structural features and properties of films and powders based on naphthalocyanines

    NASA Astrophysics Data System (ADS)

    Ramonova, A. G.; Nakusov, A. T.; Sozanov, V. G.; Bliev, A. P.; Magkoev, T. T.

    2015-06-01

    The template synthesis is used to produce powders and films based on naphthalocyanines and the corresponding metal complexes (Pc, CuPc, and NiPc). The atomic-force microscopy (AFM) and X-ray phase analysis (XPA) are employed in the study of structure and phase of fine powders and nanostructured films. The AFM data are used to determine the orientation and density of primary particles packed in the film. The XPA method is used to study the chemical composition and crystal structure of the synthesized samples. The regularities related to the structural features that affect the electrophysical properties of the films under study are revealed.

  6. AFM analysis of bleaching effects on dental enamel microtopography

    NASA Astrophysics Data System (ADS)

    Pedreira de Freitas, Ana Carolina; Espejo, Luciana Cardoso; Botta, Sergio Brossi; Teixeira, Fernanda de Sa; Luz, Maria Aparecida A. Cerqueira; Garone-Netto, Narciso; Matos, Adriana Bona; Salvadori, Maria Cecilia Barbosa da Silveira

    2010-02-01

    The purpose of this in vitro study was to test a new methodology to evaluate the effects of 35% hydrogen peroxide agent on the microtopography of sound enamel using an atomic force microscope (AFM). The buccal sound surfaces of three extracted human lower incisors were used, without polishing the surfaces to maintain them with natural morphology. These unpolished surfaces were subjected to bleaching procedure with 35% hydrogen peroxide that consisted of 4 applications of the bleaching agent on enamel surfaces for 10 min each application. Surface images were obtained in a 15 μm × 15 μm area using an AFM. The roughness (Ra and RMS) and the power spectral density (PSD) were obtained before and after the bleaching treatment. As results we could inquire that the PSD analyses were very suitable to identifying the morphological changes on the surfaces, while the Ra and RMS parameters were insufficient to represent the morphological alterations promoted by bleaching procedure on enamel. The morphological wavelength in the range of visible light spectrum (380-750 nm) was analyzed, showing a considerable increase of the PSD with the bleaching treatment.

  7. An approach towards 3D sensitive AFM cantilevers

    NASA Astrophysics Data System (ADS)

    Koops, Richard; Fokkema, Vincent

    2014-04-01

    The atomic force microscope (AFM) tapping mode is a highly sensitive local probing technique that is very useful to study and measure surface properties down to the atomic scale. The tapping mode is mostly implemented using the resonance of the first bending mode of the cantilever and therefore provides sensitivity mainly along the direction of this oscillation. Driven by the semiconductor industry, there is an increasing need for accurate measurements of nanoscale structures for side wall characterization by AFM that requires additional sensitivity in the lateral direction. The conventional tapping mode has been augmented by various authors, for example by tilting the cantilever system (Cho et al 2011 Rev. Sci. Instrum. 82 023707) to access the sidewall or using a torsion mode (Dai et al 2011 Meas. Sci. Technol. 22 094009) of the cantilever to provide additional lateral sensitivity. These approaches however trade lateral sensitivity for vertical sensitivity or still lack sensitivity in the remaining lateral direction. We present an approach towards true 3D sensitivity for AFM cantilevers based on simultaneous excitation and optical detection of multiple cantilever resonance modes along three axes. Tuning the excitation of the cantilever to specific frequencies provides a mechanism to select only those cantilever modes that have the desired characteristics. Additionally, cantilever engineering has been used to design and create a substructure within the cantilever that has been optimized for specific resonance behavior around 4 MHz. In contrast to the conventional approach of using a piezo to actuate the cantilever modulation, we present results on photo-thermal excitation using an intensity modulated low-power laser source. By tightly focusing the excitation spot on the cantilever we were able to attain a deflection efficiency of 0.7 nm µW-1 for the first bending mode. The presented approach results in an efficient all optical excitation and deflection detection

  8. Epitope mapping of monoclonal antibody HPT-101: a study combining dynamic force spectroscopy, ELISA and molecular dynamics simulations

    NASA Astrophysics Data System (ADS)

    Stangner, Tim; Angioletti-Uberti, Stefano; Knappe, Daniel; Singer, David; Wagner, Carolin; Hoffmann, Ralf; Kremer, Friedrich

    2015-12-01

    By combining enzyme-linked immunosorbent assay (ELISA) and optical tweezers-assisted dynamic force spectroscopy (DFS), we identify for the first time the binding epitope of the phosphorylation-specific monoclonal antibody (mAb) HPT-101 to the Alzheimer's disease relevant peptide tau[pThr231/pSer235] on the level of single amino acids. In particular, seven tau isoforms are synthesized by replacing binding relevant amino acids by a neutral alanine (alanine scanning). From the binding between mAb HPT-101 and the alanine-scan derivatives, we extract specific binding parameters such as bond lifetime {τ }0, binding length {x}{ts}, free energy of activation {{Δ }}G (DFS) and affinity constant {K}{{a}} (ELISA, DFS). Based on these quantities, we propose criteria to identify essential, secondary and non-essential amino acids, being representative of the antibody binding epitope. The obtained results are found to be in full accord for both experimental techniques. In order to elucidate the microscopic origin of the change in binding parameters, we perform molecular dynamics (MD) simulations of the free epitope in solution for both its parent and modified form. By taking the end-to-end distance {d}{{E}-{{E}}} and the distance between the α-carbons {d}{{C}-{{C}}} of the phosphorylated residues as gauging parameters, we measure how the structure of the epitope depends on the type of substitution. In particular, whereas {d}{{C}-{{C}}} is sometimes conserved between the parent and modified form, {d}{{E}-{{E}}} strongly changes depending on the type of substitution, correlating well with the experimental data. These results are highly significant, offering a detailed microscopic picture of molecular recognition.

  9. Tapping and contact mode imaging of native chromosomes and extraction of genomic DNA using AFM tips

    NASA Astrophysics Data System (ADS)

    Sun, Yingchun; Arakawa, Hideo; Osada, Toshiya; Ikai, Atsushi

    2002-03-01

    It is very important both in medicine and biology to clarify the chromosomal structure to understand its functions. In a standard cytogenetic procedure, chromosomes are often fixed in a mixture of acetic acid and methanol. This process most likely changes the mechanical property of chromosomes. We adopted a method to prepare native and unfixed chromosomes from mouse 3T3 cells and used tapping and contact mode atomic force microscopy (AFM) to image and manipulate them. Modified AFM tips were used to image chromosomes in contact mode in air, and then the chromosome samples were immobilized on a substrate and placed in a buffer solution to pull out DNA-histone complexes from them after they were optimally treated with trypsin. From the AFM images, we could see several bands and granular structures on chromosomes. We obtained force curves indicating long fiber extensions from native chromosomes both with low (in high concentration of NaCl) and high forces (physiological conditions). The result suggested that the degree of chromosome condensation decreased in high concentration of salt. It agrees with the known fact of histone H1 dissociation in a high concentration of salt. We intend to pull out DNA-histone complexes from chromosomes for later molecular operations on them using an AFM.

  10. Nanomechanical probing of soft matter through hydrophobic AFM tips fabricated by two-photon polymerization

    NASA Astrophysics Data System (ADS)

    Suriano, Raffaella; Zandrini, Tommaso; De Marco, Carmela; Osellame, Roberto; Turri, Stefano; Bragheri, Francesca

    2016-04-01

    Atomic force microscopy (AFM) nanoindentation of soft materials is a powerful tool for probing mechanical properties of biomaterials. Though many results have been reported in this field over the last decade, adhesion forces between the tip and the sample hinder the elastic modulus measurement when hydrophilic soft samples are investigated. Here, two-photon polymerization (2PP) technology was used to fabricate hydrophobic perfluoropolyether-based AFM tips. The hydrophobic 2PP tips allowed us to overcome the limitations of commercial and functionalized tips as well as to successfully measure the elastic modulus of medically relevant soft materials in air. Our results obtained in the characterization of poly(dimethyl siloxane) and polyethylene glycol hydrogels showed lower adhesion forces over a larger measurement range when compared to measurements performed with commercial tips. The elastic moduli measured by means of hydrophobic 2PP AFM tips were also found to be comparable to those obtained using conventional techniques for macroscopic samples. We successfully showed that the hydrophobic AFM tips developed by this highly versatile technology enable the study of mechanical properties of soft matter, benefiting from reduced sample-tip interactions, and a custom-made shape and dimension of the tips.

  11. Comparative studies of thin film growth on aluminium by AFM, TEM and GDOES characterization

    NASA Astrophysics Data System (ADS)

    Qi, Jiantao; Thompson, George E.

    2016-07-01

    In this present study, comparative studies of trivalent chromium conversion coating formation, associated with aluminium dissolution process, have been investigated using atomic force microscopy (AFM), transmission electron microscopy (TEM) and glow-discharge optical emission spectroscopy (GDOES). High-resolution electron micrographs revealed the evident and uniform coating initiation on the whole surface after conversion treatment for only 30 s, although a network of metal ridges was created by HF etching pre-treatment. In terms of conversion treatment process on electropolished aluminium, constant kinetics of coating growth, ∼0.30 ± 0.2 nm/s, were found after the prolonged conversion treatment for 600 s. The availability of electrolyte anions for coating deposition determined the growth process. Simultaneously, a proceeding process of aluminium dissolution during conversion treatment, of ∼0.11 ± 0.02 nm/s, was found for the first time, indicating constant kinetics of anodic reactions. The distinct process of aluminium consumption was assigned with loss of corrosion protection of the deposited coating material as evidenced in the electrochemical impedance spectroscopy. Based on the present data, a new mechanism of coating growth on aluminium was proposed, and it consisted of an activation period (0-30 s), a linear growth period (0.30 nm/s, up for 600 s) and limited growth period (0.17 nm/s, 600-1200 s). In addition, the air-drying post-treatment and a high-vacuum environment in the microscope revealed a coating shrinkage, especially in the coatings after conversion treatments for longer time.

  12. A rapid and automated relocation method of an AFM probe for high-resolution imaging

    NASA Astrophysics Data System (ADS)

    Zhou, Peilin; Yu, Haibo; Shi, Jialin; Jiao, Niandong; Wang, Zhidong; Wang, Yuechao; Liu, Lianqing

    2016-09-01

    The atomic force microscope (AFM) is one of the most powerful tools for high-resolution imaging and high-precision positioning for nanomanipulation. The selection of the scanning area of the AFM depends on the use of the optical microscope. However, the resolution of an optical microscope is generally no larger than 200 nm owing to wavelength limitations of visible light. Taking into consideration the two determinants of relocation—relative angular rotation and positional offset between the AFM probe and nano target—it is therefore extremely challenging to precisely relocate the AFM probe to the initial scan/manipulation area for the same nano target after the AFM probe has been replaced, or after the sample has been moved. In this paper, we investigate a rapid automated relocation method for the nano target of an AFM using a coordinate transformation. The relocation process is both simple and rapid; moreover, multiple nano targets can be relocated by only identifying a pair of reference points. It possesses a centimeter-scale location range and nano-scale precision. The main advantages of this method are that it overcomes the limitations associated with the resolution of optical microscopes, and that it is label-free on the target areas, which means that it does not require the use of special artificial markers on the target sample areas. Relocation experiments using nanospheres, DNA, SWCNTs, and nano patterns amply demonstrate the practicality and efficiency of the proposed method, which provides technical support for mass nanomanipulation and detection based on AFM for multiple nano targets that are widely distributed in a large area.

  13. A rapid and automated relocation method of an AFM probe for high-resolution imaging.

    PubMed

    Zhou, Peilin; Yu, Haibo; Shi, Jialin; Jiao, Niandong; Wang, Zhidong; Wang, Yuechao; Liu, Lianqing

    2016-09-30

    The atomic force microscope (AFM) is one of the most powerful tools for high-resolution imaging and high-precision positioning for nanomanipulation. The selection of the scanning area of the AFM depends on the use of the optical microscope. However, the resolution of an optical microscope is generally no larger than 200 nm owing to wavelength limitations of visible light. Taking into consideration the two determinants of relocation-relative angular rotation and positional offset between the AFM probe and nano target-it is therefore extremely challenging to precisely relocate the AFM probe to the initial scan/manipulation area for the same nano target after the AFM probe has been replaced, or after the sample has been moved. In this paper, we investigate a rapid automated relocation method for the nano target of an AFM using a coordinate transformation. The relocation process is both simple and rapid; moreover, multiple nano targets can be relocated by only identifying a pair of reference points. It possesses a centimeter-scale location range and nano-scale precision. The main advantages of this method are that it overcomes the limitations associated with the resolution of optical microscopes, and that it is label-free on the target areas, which means that it does not require the use of special artificial markers on the target sample areas. Relocation experiments using nanospheres, DNA, SWCNTs, and nano patterns amply demonstrate the practicality and efficiency of the proposed method, which provides technical support for mass nanomanipulation and detection based on AFM for multiple nano targets that are widely distributed in a large area. PMID:27559679

  14. Development of a 3D-AFM for true 3D measurements of nanostructures

    NASA Astrophysics Data System (ADS)

    Dai, Gaoliang; Häßler-Grohne, Wolfgang; Hüser, Dorothee; Wolff, Helmut; Danzebrink, Hans-Ulrich; Koenders, Ludger; Bosse, Harald

    2011-09-01

    The development of advanced lithography requires highly accurate 3D metrology methods for small line structures of both wafers and photomasks. Development of a new 3D atomic force microscopy (3D-AFM) with vertical and torsional oscillation modes is introduced in this paper. In its configuration, the AFM probe is oscillated using two piezo actuators driven at vertical and torsional resonance frequencies of the cantilever. In such a way, the AFM tip can probe the surface with a vertical and a lateral oscillation, offering high 3D probing sensitivity. In addition, a so-called vector approach probing (VAP) method has been applied. The sample is measured point-by-point using this method. At each probing point, the tip is approached towards the surface until the desired tip-sample interaction is detected and then immediately withdrawn from the surface. Compared to conventional AFMs, where the tip is kept continuously in interaction with the surface, the tip-sample interaction time using the VAP method is greatly reduced and consequently the tip wear is reduced. Preliminary experimental results show promising performance of the developed system. A measurement of a line structure of 800 nm height employing a super sharp AFM tip could be performed with a repeatability of its 3D profiles of better than 1 nm (p-v). A line structure of a Physikalisch-Technische Bundesanstalt photomask with a nominal width of 300 nm has been measured using a flared tip AFM probe. The repeatability of the middle CD values reaches 0.28 nm (1σ). A long-term stability investigation shows that the 3D-AFM has a high stability of better than 1 nm within 197 measurements taken over 30 h, which also confirms the very low tip wear.

  15. A software tool for STED-AFM correlative super-resolution microscopy

    NASA Astrophysics Data System (ADS)

    Koho, Sami; Deguchi, Takahiro; Löhmus, Madis; Näreoja, Tuomas; Hänninen, Pekka E.

    2015-03-01

    Multi-modal correlative microscopy allows combining the strengths of several imaging techniques to provide unique contrast. However it is not always straightforward to setup instruments for such customized experiments, as most microscope manufacturers use their own proprietary software, with limited or no capability to interface with other instruments - this makes correlation of the multi-modal data extremely challenging. We introduce a new software tool for simultaneous use of a STimulated Emission Depletion (STED) microscope with an Atomic Force Microscope (AFM). In our experiments, a Leica TCS STED commercial super-resolution microscope, together with an Agilent 5500ilm AFM microscope was used. With our software, it is possible to synchronize the data acquisition between the STED and AFM instruments, as well as to perform automatic registration of the AFM images with the super-resolution STED images. The software was realized in LabVIEW; the registration part was also implemented as an ImageJ script. The synchronization was realized by controlling simple trigger signals, also available in the commercial STED microscope, with a low-cost National Instruments USB-6501 digital I/O card. The registration was based on detecting the positions of the AFM tip inside the STED fieldof-view, which were then used as registration landmarks. The registration should work on any STED and tip-scanning AFM microscope combination, at nanometer-scale precision. Our STED-AFM correlation method has been tested with a variety of nanoparticle and fixed cell samples. The software will be released under BSD open-source license.

  16. AFM characterization of nonwoven material functionalized by ZnO sputter coating

    SciTech Connect

    Deng Bingyao; Yan Xiong; Wei Qufu Gao Weidong

    2007-10-15

    Sputter coatings provide new approaches to the surface functionalization of textile materials. In this study, polyethylene terephthalate (PET) nonwoven material was used as a substrate for creating functional nanostructures on the fiber surfaces. A magnetron sputter coating was used to deposit functional zinc oxide (ZnO) nanostructures onto the nonwoven substrate. The evolution of the surface morphology of the fibers in the nonwoven web was examined using atomic force microscopy (AFM). The AFM observations revealed a significant difference in the morphology of the fibers before and after the sputter coating. The AFM images also indicated the effect of the sputtering conditions on the surface morphology of the fibers. The increase in the sputtering time led to the growth of the ZnO grains on the fiber surfaces. The higher pressure in the sputtering chamber could cause the formation of larger grains on the fiber surfaces. The higher power used also generated larger grains on the fiber surfaces.

  17. Crystallization of Probucol in Nanoparticles Revealed by AFM Analysis in Aqueous Solution.

    PubMed

    Egami, Kiichi; Higashi, Kenjirou; Yamamoto, Keiji; Moribe, Kunikazu

    2015-08-01

    The crystallization behavior of a pharmaceutical drug in nanoparticles was directly evaluated by atomic force microscopy (AFM) force curve measurements in aqueous solution. A ternary spray-dried sample (SPD) was prepared by spray drying the organic solvent containing probucol (PBC), hypromellose (HPMC), and sodium dodecyl sulfate (SDS). The amorphization of PBC in the ternary SPD was confirmed by powder X-ray diffraction (PXRD) and solid-state 13C NMR measurements. A nanosuspension containing quite small particles of 25 nm in size was successfully prepared immediately after dispersion of the ternary SPD into water. Furthermore, solution-state 1H NMR measurements revealed that a portion of HPMC coexisted with PBC as a mixed state in the freshly prepared nanosuspension particles. After storing the nanosuspension at 25 °C, a gradual increase in the size of the nanoparticles was observed, and the particle size changed to 93.9 nm after 7 days. AFM enabled the direct observation of the morphology and agglomeration behavior of the nanoparticles in water. Moreover, AFM force-distance curves were changed from (I) to (IV), depending on the storage period, as follows: (I) complete indentation within an applied force of 1 nN, (II) complete indentation with an applied force of 1-5 nN, (III) partial indentation with an applied force of 5 nN, and (IV) nearly no indentation with an applied force of 5 nN. This stiffness increase of the nanoparticles was attributed to gradual changes in the molecular state of PBC from the amorphous to the crystal state. Solid-state 13C NMR measurements of the freeze-dried samples demonstrated the presence of metastable PBC Form II crystals in the stored nanosuspension, strongly supporting the AFM results.

  18. Quantitative Nano-structural and Single Molecule Force Spectroscopy bio-molecular analysis of human saliva derived exosomes

    PubMed Central

    Sharma, Shivani; Gillespie, Boyd M; Palanisamy, Viswanathan; Gimzewski, James K.

    2011-01-01

    Exosomes are naturally occurring nanoparticles with unique structure, surface biochemistry and mechanical characteristics. These distinct nanometer sized bio-particles are secreted from the surface of oral epithelial cells into saliva, and are of interest as oral-cancer biomarkers. We use high- resolution AFM to show single vesicle quantitative differences between exosomes derived from normal and oral cancer patient’s saliva. Compared to normal exosomes (circular; 67.4 ± 2.9 nm), our findings indicate that cancer exosomes populations are significantly increased in saliva and display irregular morphologies, increased vesicle size (98.3 ± 4.6 nm) and higher inter-vesicular aggregation. At the single vesicle level, cancer exosomes exhibit significantly (P<0.05) increased CD63 surface densities. To our knowledge, it represents the first report detecting single exosome surface protein variations. Additionally, high- resolution AFM imaging of cancer saliva samples revealed discrete multi-vesicular bodies with intra-luminal exosomes enclosed. We discuss the use of quantitative, nanoscale ultra-structural and surface bio-molecular analysis of saliva exosomes, at the single vesicle and single protein level sensitivity, as a potentially new oral cancer diagnostic. PMID:22017459

  19. AFM Bio-Mechanical Investigation of the Taxol Treatment of Breast Cancer Cells

    NASA Astrophysics Data System (ADS)

    Smith, Dylan; Patel, Dipika; Monjaraz, Fernando; Park, Soyeun

    2009-10-01

    Cancerous cells are known to be softer and easier to deform than normal cells. Changes in mechanical properties originate from the alteration of the actin cytoskeleton. The mechanism of cancer treatment using Taxol is related to the stabilization of microtubules. It has been shown that Taxol binds to polymerized tublin, stabilizes it against disassembly, and consequently inhibits cell division. An accurate quantitative study still lacks to relate the microtubule stabilizing effect with the cellular mechanical properties. We utilized our AFM to study changes in elastic properties of treated breast cancer cells. The AFM has several advantages for precise force measurements on a localized region with nanometer lateral dimension. In previous AFM studies, measurable contributions from the underlying hard substrate have been an obstacle to accurately determine the properties on thin samples. We modified our AFM tip to obtain the exact deformation profile as well as reducing the high stresses produced. We have probed depth profiles of mechanical properties of the taxol-treated and untreated cells by varying the indentation depth of the AFM-nanoindenting experiments.

  20. Fractal analysis of AFM images of the surface of Bowman's membrane of the human cornea.

    PubMed

    Ţălu, Ştefan; Stach, Sebastian; Sueiras, Vivian; Ziebarth, Noël Marysa

    2015-04-01

    The objective of this study is to further investigate the ultrastructural details of the surface of Bowman's membrane of the human cornea, using atomic force microscopy (AFM) images. One representative image acquired of Bowman's membrane of a human cornea was investigated. The three-dimensional (3-D) surface of the sample was imaged using AFM in contact mode, while the sample was completely submerged in optisol solution. Height and deflection images were acquired at multiple scan lengths using the MFP-3D AFM system software (Asylum Research, Santa Barbara, CA), based in IGOR Pro (WaveMetrics, Lake Oswego, OR). A novel approach, based on computational algorithms for fractal analysis of surfaces applied for AFM data, was utilized to analyze the surface structure. The surfaces revealed a fractal structure at the nanometer scale. The fractal dimension, D, provided quantitative values that characterize the scale properties of surface geometry. Detailed characterization of the surface topography was obtained using statistical parameters, in accordance with ISO 25178-2: 2012. Results obtained by fractal analysis confirm the relationship between the value of the fractal dimension and the statistical surface roughness parameters. The surface structure of Bowman's membrane of the human cornea is complex. The analyzed AFM images confirm a fractal nature of the surface, which is not taken into account by classical surface statistical parameters. Surface fractal dimension could be useful in ophthalmology to quantify corneal architectural changes associated with different disease states to further our understanding of disease evolution.

  1. Submolecular Resolution Imaging of Molecules by Atomic Force Microscopy: The Influence of the Electrostatic Force.

    PubMed

    van der Lit, Joost; Di Cicco, Francesca; Hapala, Prokop; Jelinek, Pavel; Swart, Ingmar

    2016-03-01

    The forces governing the contrast in submolecular resolution imaging of molecules with atomic force microscopy (AFM) have recently become a topic of intense debate. Here, we show that the electrostatic force is essential to understand the contrast in atomically resolved AFM images of polar molecules. Specifically, we image strongly polarized molecules with negatively and positively charged tips. A contrast inversion is observed above the polar groups. By taking into account the electrostatic forces between tip and molecule, the observed contrast differences can be reproduced using a molecular mechanics model. In addition, we analyze the height dependence of the various force components contributing to the high-resolution AFM contrast.

  2. Submolecular Resolution Imaging of Molecules by Atomic Force Microscopy: The Influence of the Electrostatic Force

    NASA Astrophysics Data System (ADS)

    van der Lit, Joost; Di Cicco, Francesca; Hapala, Prokop; Jelinek, Pavel; Swart, Ingmar

    2016-03-01

    The forces governing the contrast in submolecular resolution imaging of molecules with atomic force microscopy (AFM) have recently become a topic of intense debate. Here, we show that the electrostatic force is essential to understand the contrast in atomically resolved AFM images of polar molecules. Specifically, we image strongly polarized molecules with negatively and positively charged tips. A contrast inversion is observed above the polar groups. By taking into account the electrostatic forces between tip and molecule, the observed contrast differences can be reproduced using a molecular mechanics model. In addition, we analyze the height dependence of the various force components contributing to the high-resolution AFM contrast.

  3. Studying Chemical Reactions, One Bond at a Time, with Single Molecule AFM Techniques

    NASA Astrophysics Data System (ADS)

    Fernandez, Julio M.

    2008-03-01

    The mechanisms by which mechanical forces regulate the kinetics of a chemical reaction are unknown. In my lecture I will demonstrate how we use single molecule force-clamp spectroscopy and protein engineering to study the effect of force on the kinetics of thiol/disulfide exchange. Reduction of disulfide bond via the thiol/disulfide exchange chemical reaction is crucial in regulating protein function and is of common occurrence in mechanically stressed proteins. While reduction is thought to proceed through a substitution nucleophilic bimolecular (SN2) reaction, the role of a mechanical force in modulating this chemical reaction is unknown. We apply a constant stretching force to single engineered disulfide bonds and measure their rate of reduction by dithiothreitol (DTT). We find that while the reduction rate is linearly dependent on the concentration of DTT, it is exponentially dependent on the applied force, increasing 10-fold over a 300 pN range. This result predicts that the disulfide bond lengthens by 0.34 å at the transition state of the thiol/disulfide exchange reaction. In addition to DTT, we also study the reduction of the engineered disulfide bond by the E. coli enzyme thioredoxin (Trx). Thioredoxins are enzymes that catalyze disulfide bond reduction in all organisms. As before, we apply a mechanical force in the range of 25-450 pN to the engineered disulfide bond substrate and monitor the reduction of these bonds by individual enzymes. In sharp contrast with the data obtained with DTT, we now observe two alternative forms of the catalytic reaction, the first requiring a reorientation of the substrate disulfide bond, causing a shortening of the substrate polypeptide by 0.76±0.07 å, and the second elongating the substrate disulfide bond by 0.21±0.01 å. These results support the view that the Trx active site regulates the geometry of the participating sulfur atoms, with sub-ångström precision, in order to achieve efficient catalysis. Single molecule

  4. Mechanical properties study of SW480 cells based on AFM.

    PubMed

    Liu, Xiaogang; Song, Zhengxun; Qu, Yingmin; Wang, Guoliang; Wang, Zuobin

    2015-08-01

    Since the invention of the atomic force microscope (AFM), it has been widely applied in biomedicine. One of the most important applications is used as an indenter tool to do the indentation experiment in order to get the mechanical properties of cells. In this paper, SW480 cells were used as the test subjects. Through the analysis of the contact and indentation, Young's modulus (E), which is an important parameter of cancer cells, has been estimated. Experimental results show that different mechanical models should be chosen to calculate the E in different indentation depths. Here, the E of SW480 cells was (2.5 ± 0.8) KPa at the indentation depth of 99 nm.

  5. Mechanical Characterization of Photo-crosslinkable Hydrogels with AFM

    NASA Astrophysics Data System (ADS)

    McKenna, Alyssa; Byun, Myunghwan; Hayward, Ryan; Aidala, Katherine

    2012-02-01

    Stimuli-responsive hydrogel films formed from photo-crosslinkable polymers are versatile materials for controlled drug delivery devices, three-dimensional micro-assemblies, and components in microfluidic systems. For such applications, it is important to understand both the mechanical properties and the dynamics responses of these materials. We describe the use of atomic force microscope (AFM) based indentation experiments to characterize the properties of poly(N-isopropylacrylamide) copolymer films, crosslinked by activation of pendent benzophenone units using ultraviolet light. In particular, we study how the elastic modulus of the material, determined using the Johnson, Kendall, and Roberts model, depends on UV dose, and simultaneously investigate stress relaxation in these materials in the context of viscoelastic and poroelastic relaxation models.

  6. AFM of self-assembled lambda DNA-histone networks.

    PubMed

    Liu, YuYing; Guthold, Martin; Snyder, Matthew J; Lu, HongFeng

    2015-10-01

    Atomic force microscopy (AFM) was used to investigate the self-assembly behavior of λ-DNA and histones at varying histone:DNA ratios. Without histones and at the lowest histone:DNA ratio (less than one histone per 1000 base pairs of DNA), the DNA appeared as individual (uncomplexed), double-stranded DNA molecules. At increasing histone concentrations (one histone per 500, 250 and 167 base pairs of DNA), the DNA molecules started to form extensive polygonal networks of mostly pentagons and hexagons. The observed networks might be one of the naturally occurring, stable DNA-histone structures. The condensing effects of the divalent cations Mg(2+) and Ca(2+) on the DNA-histone complexes were also investigated. The networks persisted at high Mg(2+) concentration (20mM) and the highest histone concentration. At high Ca(2+) concentration and the highest histone concentration, the polygonal network disappeared and, instead, individual, tightly condensed aggregates were formed. PMID:26141439

  7. Accurate and precise calibration of AFM cantilever spring constants using laser Doppler vibrometry.

    PubMed

    Gates, Richard S; Pratt, Jon R

    2012-09-21

    Accurate cantilever spring constants are important in atomic force microscopy both in control of sensitive imaging and to provide correct nanomechanical property measurements. Conventional atomic force microscope (AFM) spring constant calibration techniques are usually performed in an AFM. They rely on significant handling and often require touching the cantilever probe tip to a surface to calibrate the optical lever sensitivity of the configuration. This can damage the tip. The thermal calibration technique developed for laser Doppler vibrometry (LDV) can be used to calibrate cantilevers without handling or touching the tip to a surface. Both flexural and torsional spring constants can be measured. Using both Euler-Bernoulli modeling and an SI traceable electrostatic force balance technique as a comparison we demonstrate that the LDV thermal technique is capable of providing rapid calibrations with a combination of ease, accuracy and precision beyond anything previously available.

  8. Molecular Dynamic Simulations of Interaction of an AFM Probe with the Surface of an SCN Sample

    NASA Technical Reports Server (NTRS)

    Bune, Adris; Kaukler, William; Rose, M. Franklin (Technical Monitor)

    2001-01-01

    Molecular dynamic (MD) simulations is conducted in order to estimate forces of probe-substrate interaction in the Atomic Force Microscope (AFM). First a review of available molecular dynamic techniques is given. Implementation of MD simulation is based on an object-oriented code developed at the University of Delft. Modeling of the sample material - succinonitrile (SCN) - is based on the Lennard-Jones potentials. For the polystyrene probe an atomic interaction potential is used. Due to object-oriented structure of the code modification of an atomic interaction potential is straight forward. Calculation of melting temperature is used for validation of the code and of the interaction potentials. Various fitting parameters of the probe-substrate interaction potentials are considered, as potentials fitted to certain properties and temperature ranges may not be reliable for the others. This research provides theoretical foundation for an interpretation of actual measurements of an interaction forces using AFM.

  9. Switching surface polarization of atomic force microscopy probe utilizing photoisomerization of photochromic molecules

    SciTech Connect

    Aburaya, Yoshihiro; Nomura, Hikaru; Kageshima, Masami; Naitoh, Yoshitaka; Li, Yan Jun; Sugawara, Yasuhiro

    2011-03-15

    An attempt to develop an atomic force microscopy (AFM) probe with optically switchable polarization is described. Modification with a single molecular layer of photochromic molecules was attempted onto a Si substrate that is a prototype for a probe surface. Polarization switching caused by alternate irradiation of UV and visible lights were detected using the electrostatic force?>spectroscopy (EFS) technique. Si substrates modified with spiropyran and azobenzene exhibited reversible polarization switching that caused changes in CPD of about 100 and 50 mV, respectively. Modification with spiropyran was also attempted onto a Si probe and resulted in a CPD change of about 100 mV. It was confirmed that modification of an AFM probe or substrate with a single molecular layer of photochromic molecules can generate surface polarization switching of a mechanically detectable level.

  10. Force spectroscopy and fluorescence microscopy of dsDNA–YOYO-1 complexes: implications for the structure of dsDNA in the overstretching region

    PubMed Central

    Murade, Chandrashekhar U.; Subramaniam, Vinod; Otto, Cees; Bennink, Martin L.

    2010-01-01

    When individual dsDNA molecules are stretched beyond their B-form contour length, they reveal a structural transition in which the molecule extends 1.7 times its contour length. The nature of this transition is still a subject of debate. In the first model, the DNA helix unwinds and combined with the tilting of the base pairs (which remain intact), results in a stretched form of DNA (also known as S-DNA). In the second model the base pairs break resulting effectively in two single-strands, which is referred to as force-induced melting. Here a combination of optical tweezers force spectroscopy with fluorescence microscopy was used to study the structure of dsDNA in the overstretching regime. When dsDNA was stretched in the presence of 10 nM YOYO-1 an initial increase in total fluorescence intensity of the dye–DNA complex was observed and at an extension where the dsDNA started to overstretch the fluorescence intensity leveled off and ultimately decreased when stretched further into the overstretching region. Simultaneous force spectroscopy and fluorescence polarization microscopy revealed that the orientation of dye molecules did not change significantly in the overstretching region (78.0°± 3.2°). These results presented here clearly suggest that, the structure of overstretched dsDNA can be explained accurately by force induced melting. PMID:20129944

  11. Imaging and measuring the biophysical properties of Fc gamma receptors on single macrophages using atomic force microscopy

    SciTech Connect

    Li, Mi; Liu, Lianqing; Xi, Ning; Wang, Yuechao; Xiao, Xiubin; Zhang, Weijing

    2013-09-06

    Highlights: •Nanoscale cellular ultra-structures of macrophages were observed. •The binding affinities of FcγRs were measured directly on macrophages. •The nanoscale distributions of FcγRs were mapped on macrophages. -- Abstract: Fc gamma receptors (FcγR), widely expressed on effector cells (e.g., NK cells, macrophages), play an important role in clinical cancer immunotherapy. The binding of FcγRs to the Fc portions of antibodies that are attached to the target cells can activate the antibody-dependent cell-mediated cytotoxicity (ADCC) killing mechanism which leads to the lysis of target cells. In this work, we used atomic force microscopy (AFM) to observe the cellular ultra-structures and measure the biophysical properties (affinity and distribution) of FcγRs on single macrophages in aqueous environments. AFM imaging was used to obtain the topographies of macrophages, revealing the nanoscale cellular fine structures. For molecular interaction recognition, antibody molecules were attached onto AFM tips via a heterobifunctional polyethylene glycol (PEG) crosslinker. With AFM single-molecule force spectroscopy, the binding affinities of FcγRs were quantitatively measured on single macrophages. Adhesion force mapping method was used to localize the FcγRs, revealing the nanoscale distribution of FcγRs on local areas of macrophages. The experimental results can improve our understanding of FcγRs on macrophages; the established approach will facilitate further research on physiological activities involved in antibody-based immunotherapy.

  12. Controlled AFM detachments and movement of nanoparticles: gold clusters on HOPG at different temperatures.

    PubMed

    Tripathi, Manoj; Paolicelli, Guido; D'Addato, Sergio; Valeri, Sergio

    2012-06-22

    The effect of temperature on the onset of movement of gold nanoclusters (diameter 27 nm) deposited on highly oriented pyrolytic graphite (HOPG) has been studied by atomic force microscopy (AFM) techniques. Using the AFM with amplitude modulation (tapping mode AFM) we have stimulated and controlled the movement of individual clusters. We show how, at room temperature, controlled detachments and smooth movements can be obtained for clusters having dimensions comparable to or smaller than the tip radius. Displacement is practically visible in real time and it can be started and stopped easily by adjusting only one parameter, the tip amplitude oscillation. Analysing the energy dissipation signal at the onset of nanocluster sliding we evaluated a detachment threshold energy as a function of temperature in the range 300-413 K. We also analysed single cluster thermal induced displacement and combining this delicate procedure with AFM forced movement behaviour we conclude that detachment threshold energy is directly related to the activation energy of nanocluster diffusion and it scales linearly with temperature as expected for a single-particle thermally activated process.

  13. Difference in cellular mechanics of cancer and normal cervical cells as seen with the AFM

    NASA Astrophysics Data System (ADS)

    Sokolov, Igor; Iyer, Swaminathan; Subba-Rao, Venkatesh; Woodworth, Craig

    2006-03-01

    Oncogenically transformed cells differ from their normal counter parts in many aspects, including organization and the amount of cytoskeleton. Consequently it is natural to expect to see the difference in cellular mechanics. Here we will present the study of such differences by using atomic force microscopy (AFM) in-vitro. So far the present research is the first study of mechanics of cervical cells, and the third comparative study of differences between mechanics of cancer and normal cells down with the help of AFM. Using a micron size silica ball as the AFM probe, we presumably do not overstress the cell surface as it can be in the case of the sharp AFM tip, and consequently, we may use the classical Hertz model. In contrast to the reported previously studies (bladder and fibroblast cells), we found that oncogenically transformed cervical cells are more rigid than the normal cells. The reason for such difference will be discussed. To demonstrate the complexity of the problem, we study cell mechanics in detail. The Young's modulus of rigidity clearly shows two separate regions of rigidity depending on the depth of the probe penetration. There may be two alternative explanations of the difference in rigidity of this top layer: it is either the cell membrane layer or detected long-range (presumably steric) forces due to the molecular ``brush'' of glycocalyx molecules. Experiments and modal calculations will be presented to choose between these two possibilities.

  14. Direct measurement of single-molecule visco-elasticity in atomic force microscope force-extension experiments.

    PubMed

    Bippes, Christian A; Humphris, Andrew D L; Stark, Martin; Müller, Daniel J; Janovjak, Harald

    2006-02-01

    Measuring the visco-elastic properties of biological macromolecules constitutes an important step towards the understanding of dynamic biological processes, such as cell adhesion, muscle function, or plant cell wall stability. Force spectroscopy techniques based on the atomic force microscope (AFM) are increasingly used to study the complex visco-elastic response of (bio-)molecules on a single-molecule level. These experiments either require that the AFM cantilever is actively oscillated or that the molecule is clamped at constant force to monitor thermal cantilever motion. Here we demonstrate that the visco-elasticity of single bio-molecules can readily be extracted from the Brownian cantilever motion during conventional force-extension measurements. It is shown that the characteristics of the cantilever determine the signal-to-noise (S/N) ratio and time resolution. Using a small cantilever, the visco-elastic properties of single dextran molecules were resolved with a time resolution of 8.3 ms. The presented approach can be directly applied to probe the dynamic response of complex bio-molecular systems or proteins in force-extension experiments. PMID:16237549

  15. BOREAS AFM-6 Surface Meteorological Data

    NASA Technical Reports Server (NTRS)

    Wilczak, James; Hall, Forrest G. (Editor); Newcomer, Jeffrey A. (Editor); Smith, David E. (Technical Monitor)

    2000-01-01

    The Boreal Ecosystem-Atmosphere Study (BOREAS) Airborne Fluxes and Meteorology (AFM)-6 team from the National Oceanic and Atmospheric Adminsitration/Environment Technology Laboratory (NOAA/ETL) collected surface meteorological data from 21 May to 20 Sep 1994 near the Southern Study Area-Old Jack Pine (SSA-OJP) tower site. The data are in tabular ASCII files. The surface meteorological data are available from the Earth Observing System Data and Information System (EOSDIS) Oak Ridge National Laboratory (ORNL) Distributed Active Archive Center (DAAC). The data files are available on a CD-ROM (see document number 20010000884).

  16. Nanoscale fabrication and characterization of chemically modified silicon surfaces using conductive atomic force microscopy in liquids

    NASA Astrophysics Data System (ADS)

    Kinser, Christopher Reagan

    This dissertation examines the modification and characterization of hydrogen-terminated silicon surfaces in organic liquids. Conductive atomic force microscope (cAFM) lithography is used to fabricate structures with sub-100 nm line width on H:Si(111) in n-alkanes, 1-alkenes, and 1-alkanes. Nanopatterning is accomplished by applying a positive (n-alkanes and 1-alkenes) or a negative (1-alkanes) voltage pulse to the silicon substrate with the cAFM tip connected to ground. The chemical and kinetic behavior of the patterned features is characterized using AFM, lateral force microscopy, time-of-flight secondary ion mass spectroscopy (TOF SIMS), and chemical etching. Features patterned in hexadecane, 1-octadecene, and undecylenic acid methyl ester exhibited chemical and kinetic behavior consistent with AFM field induced oxidation. The oxide features are formed due to capillary condensation of a water meniscus at the AFM tip-sample junction. A space-charge limited growth model is proposed to explain the observed growth kinetics. Surface modifications produced in the presence of neat 1-dodecyne and 1-octadecyne exhibited a reduced lateral force compared to the background H:Si(111) substrate and were resistant to a hydrofluoric acid etch, characteristics which indicate that the patterned features are not due to field induced oxidation and which are consistent with the presence of the methyl-terminated 1-alkyne bound directly to the silicon surface through silicon-carbon bonds. In addition to the cAFM patterned surfaces, full monolayers of undecylenic acid methyl ester (SAM-1) and undec-10-enoic acid 2-bromoethyl ester (SAM-2) were grown on H:Si(111) substrates using ultraviolet light. The structure and chemistry of the monolayers were characterized using AFM, TOF SIMS, X-ray photoelectron spectroscopy (XPS), X-ray reflectivity (XRR), X-ray standing waves (XSW), and X-ray fluorescence (XRF). These combined analyses provide evidence that SAM-1 and SAM-2 form dense monolayers

  17. Dynamic modeling and sensitivity analysis of dAFM in the transient and steady state motions.

    PubMed

    Payam, Amir Farokh

    2016-10-01

    In this paper, based on the slow time varying function theory, dynamical equations for the amplitude and phase of the dynamic atomic force microscope are derived. Then, the sensitivity of the amplitude and phase to the dissipative and conservative parts of interaction force is investigated. The most advantage of this dynamical model is the ability to simulate and analysis the dynamics behavior of amplitude and phase of the AFM tip motion not only in the steady state but also in the transient regime. Using numerical analysis the transient and steady state behavior of amplitude and phase is studied and the sensitivity of amplitude and phase to the interaction force is analyzed. PMID:27448201

  18. Molecular recognition force spectroscopy study of the dynamic interaction between aptamer GBI-10 and extracellular matrix protein tenascin-C on human glioblastoma cell.

    PubMed

    Li, Yongjun; Qiao, Haiyan; Yan, Wei; Zhang, Jing; Xing, Chunyan; Wang, Hongda; Zhang, Bailin; Tang, Jilin

    2013-01-01

    Molecular recognition force spectroscopy (MR-FS) was applied to investigate the dynamic interaction between aptamer GBI-10 and tenascin-C (TN-C) on human glioblastoma cell surface at single-molecule level. The unbinding force between aptamer GBI-10 and TN-C was 39 pN at the loading rate of 0.3 nN sec⁻¹. A series of kinetic parameters concerning interaction process such as the unbinding force f(u) , the association rate constant k(on) , dissociation rate constant at zero force k(off) , and dissociation constant K(D) for aptamer GBI-10/TN-C complexes were acquired. In addition, the interaction of aptamer GBI-10 with TN-C depended on the presence of Mg²⁺. This work demonstrates that MR-FS can be used as an attractive tool for exploring the interaction forces and dynamic process of aptamer and ligand at the single-molecule level. As a future perspective, MR-FS may be used as a potential diagnostic and therapeutic tool by combining with other techniques.

  19. AFM studies of semicrystalline polymer/inorganic nanocomposites

    NASA Astrophysics Data System (ADS)

    Strawhecker, Kenneth E.

    2002-01-01

    The aims of this work are to elucidate the effects of interaction strength on the crystallization of a polymer near an inorganic surface; connect filler induced polymer crystallinity with resulting property changes, especially for strongly interacting (i.e. hydrogen bonding) systems; and to devise atomic force microscopy (AFM) methods for probing the crystallinity and properties of polymer/layered silicate systems at the nanometer level. Three inorganically filled systems were studied: (1) poly(vinyl alcohol) (PVA), (2) poly(ethylene oxide) (PEO), and (3) polypropylene (PP). Since it has the strongest, interactions, the PVA system is investigated first. AFM is used in conjunction with x-ray diffraction and differential scanning calorimetry (DSC) to show that strong polymer/filler interactions can promote a different crystalline structure and a different morphology than those seen in the bulk. The study then proceeds to the weakly interacting PEO/inorganic system where it is found that the inorganic layers disrupt crystalline morphology, but do not change the crystal structure. Furthermore, crystallization always occurs in volumes away from the inorganic filler. The third system (neutral interactions), PP/inorganic is then discussed. The three systems are compared with each other, and is found that the crystalline morphology and structure is highly dependent upon the strength of interaction between the polymer and filler. Due to its far-reaching morphology changes, the strongly interacting system was chosen for property studies. The composite structure study revealed a coexistence of exfoliated and intercalated MMT layers, especially for low and moderate silicate loadings. The inorganic layers promote a new crystalline phase different than the one of the respective neat PVA, characterized by higher melting temperature and a different crystal structure. This new crystal phase reflects on the composite materials properties, which have mechanical, thermal, and water

  20. Specificity and mechanism of action of alpha-helical membrane-active peptides interacting with model and biological membranes by single-molecule force spectroscopy.

    PubMed

    Sun, Shiyu; Zhao, Guangxu; Huang, Yibing; Cai, Mingjun; Shan, Yuping; Wang, Hongda; Chen, Yuxin

    2016-01-01

    In this study, to systematically investigate the targeting specificity of membrane-active peptides on different types of cell membranes, we evaluated the effects of peptides on different large unilamellar vesicles mimicking prokaryotic, normal eukaryotic, and cancer cell membranes by single-molecule force spectroscopy and spectrum technology. We revealed that cationic membrane-active peptides can exclusively target negatively charged prokaryotic and cancer cell model membranes rather than normal eukaryotic cell model membranes. Using Acholeplasma laidlawii, 3T3-L1, and HeLa cells to represent prokaryotic cells, normal eukaryotic cells, and cancer cells in atomic force microscopy experiments, respectively, we further studied that the single-molecule targeting interaction between peptides and biological membranes. Antimicrobial and anticancer activities of peptides exhibited strong correlations with the interaction probability determined by single-molecule force spectroscopy, which illustrates strong correlations of peptide biological activities and peptide hydrophobicity and charge. Peptide specificity significantly depends on the lipid compositions of different cell membranes, which validates the de novo design of peptide therapeutics against bacteria and cancers. PMID:27363513