Parameters affecting the adhesion strength between a living cell and a colloid probe when measured by the atomic force microscope

In this study, we used the colloid probe atomic force microscopy (AFM) technique to investigate the adhesion force between a living cell and a silica colloid particle in a Leibovitz's L-15 medium (L-15). The L-15 liquid maintained the pharmaceutical conditions necessary to keep the cells alive in the outside environment during the AFM experiment. The force curves in such a system showed a steric repulsion in the compression force curve, due to the compression of the cells by the colloid probe, and an adhesion force in the decompression force curve, due to binding events between the cell and the probe. We also investigated for the first time how the position on the cell surface, the strength of the pushing force, and the residence time of the probe at the cell surface individually affected the adhesion force between a living cell and a 6.84 μm diameter silica colloid particle in L-15. The position of measuring the force on the cell surface was seen not to affect the value of the maximum adhesion force. The loading force was also seen not to notably affect the value of the maximum adhesion force, if it was small enough not to pierce and damage the cell. The residence time of the probe at the cell surface, however, clearly affected the adhesion force, where a longer residence time gave a larger maximum force. From these results, we could conclude that the AFM force measurements should be made using a loading force small enough not to damage the cell and a fixed residence time, when comparing results of different systems.     

Interfacial mechanisms governing cyclopentane clathrate hydrate adhesion/cohesion

The present work uses a micromechanical force apparatus to directly measure cyclopentane clathrate hydrate cohesive force and hydrate-steel adhesive force, as a function of contact time, contact force and temperature. We present a hydrate interparticle force model, which includes capillary and sintering contributions and is based on fundamental interparticle force theories. In this process, we estimate the cyclopentane hydrate tensile strength to be approximately 0.91 MPa. This hydrate interparticle force model also predicts the effect of temperature on hydrate particle cohesion force. Finally, we present the first direct measurements of hydrate cohesive force in the gas phase to be 9.1 ± 2.1 mN/m at approximately 3 °C (as opposed to 4.3 ± 0.4 mN/m in liquid cyclopentane).     

Force measurements for membrane protein manipulation

The force curve measurement mode of the atomic force microscope (AFM) enables us to measure hitherto unobservable mechanical properties of nanometer sized biological specimens. By applying this mode, we attempted to conduct such mechanical manipulations of membrane proteins as: (1) measurement of the separation force between a membrane bound receptor and a covalently cross-linked ligand molecule on the AFM tip; and (2) extraction of membrane proteins after harnessing them on a modified tip with covalent cross-linkers. Since the limiting tensile force of the covalent system used in our experiment was a crucial factor for successful manipulations, we first estimated the force to terminate the covalent cross-linking system at the single molecular level to be 1.6–1.7 nN, based on our previous data. The method was then applied to measure the force required to separate α₂-macroglobulin (α₂-M) from its receptor on the cell membrane using an AFM tip coated with the receptor binding form of the protein. From a bimodal distribution of rupture force, we obtained an average value of 120 pN as the force to separate a non-covalent association of α₂-M with its receptor. When modified tips with covalent cross-linkers aimed at amino groups on the cell surface were used, distribution of the rupture force shifted toward higher values, with a peak in the histogram ≈400–500 pN. Since the force to sever covalent cross-linking system was 1.6–1.7 nN, the observed force was ascribed to the force required to extract membrane proteins from the cell membrane after covalent bond formation.     

Stick slip contact mechanics between dissimilar materials: effect of charging and large friction

Measurements of the contact radius as a function of applied force between a mica surface and a silica surface (mica/silica) in air are reported. The load/unload results show that the contact radius generally increases with applied force. Because of the presence of charging due to contact electrification, both a short-range van der Waals adhesion force and longer-range electrostatic adhesive interaction contribute to the measured force. The results indicate that approximately 20% of the pull-off force is due to van der Waals forces. The contact radius versus applied force results can be fit to Johnson-Kendall-Roberts (JKR) theory by considering that only the short-range van der Waals forces contribute to the work of adhesion and subtracting a constant longer-range electrostatic force. Also, an additional and unexpected step function is superimposed on the contact radius versus applied force curve. Thus, the contact diameter increases in a stepped dependence with increasing force. The stepped contact behavior is seen only for increasing force and is not observed when symmetric mica/mica or silica/silica contacts are measured. In humid conditions, the contact diameter of the mica/silica contact increases monotonically with applied force. Friction forces between the surfaces are also measured and the shear stress of a mica/silica interface is 100 times greater than the shear stress of a mica/mica interface. This large shear stress retards the increase in contact area as the force is increased and leads to the observed stepped contact mechanics behavior.     

Ab initio computation of force constants: Part VI. Applications of the force relaxation method for geometry optimization

Techniques to improve the computational efficiency of the force relaxation method are discussed. Force constants for fragments in previously computed smaller molecules can be transferred to construct a guess force constant matrix. Additional force constants that may be needed can be computed by a procedure which uses only one additional force calculation per diagonal force constant required. A scaling technique to improve convergence on the optimized geometry is discussed.     

Dynamic force spectroscopy on soft molecular systems: Improved analysis of unbinding spectra with varying linker compliance

Dynamic force spectroscopy makes it possible to measure the breaking of single molecular bonds or the unfolding of single proteins subjected to a time-dependent pulling force. The force needed to break a single bond or to unfold a domain in a protein depends critically on the time dependence of the applied force. In this way the elastic response couples to the unbinding force. We have performed an experimental and theoretical examination of this coupling by studying the well-known biotin–streptavidin bond in systems incorporating two common types of linkers. In the first case biotin is linked by bovine serum albumin (BSA) and it is observed that this linker has a linear elastic response. More surprisingly we find that its force constant varies significantly between repeated force curves. It is demonstrated that by sorting the force curves according to the force constant of the linker we can improve the data analysis and obtain a better agreement between experimental data and theory. In the second case biotin is linked by poly(ethylene glycol) (PEG), which has a soft nonlinear elastic response. A numerical calculation of the unbinding statistics for the polymer system agrees quantitatively with experiments. It demonstrates a clear decrease in unbinding forces resulting from the polymer linker.     

A force field for some weakly coupled conjugated systems

A force field developed in a series of overlay calculations for some weakly coupled conjugated systems is discussed. The force fieid is compared with other, mainly overlay force fields of related molecules, as this gives further evidence for the transferability of the force constants. In order to be of any use for large molecules the force field must be easy to transfer to related, more complicated molecules. The disadvantages of using individual force fields are also discussed.     

Treatment of redundancies among internal coordinates in optimizing molecular mechanics force constants

The use of redundant coordinate bases in the construction of molecular mechanics force fields is discussed. It is shown that the intrinsic indeterminacy in a force field in redundant coordinates in general stems from the squares of the first-order redundancy relations. The necessity to use constraints in such a force field is pointed out, and a method to check whether or not a set of constraints makes the force field determinate is described. It is also explained how force fields corresponding to different sets of constraints can be transformed into one another. To facilitate the utilization of ab initio or other spectroscopic force fields, a procedure is given by which force constants pertaining to a nonredundant coordinate basis can be optimized in molecular mechanics calculations where redundant coordinates are used. © 1992 by John Wiley & Sons, Inc.     

Direct measurement of hydrophobic particle–bubble interactions in aqueous solutions by atomic force microscopy: Effect of particle hydrophobicity

Direct measurements of the interaction forces between a spherical silica particle and a small air bubble have been conducted in aqueous electrolyte solutions by using an atomic force microscope (AFM). The silica particle was hydrophobized with a silanating reagent, and the interaction forces were measured by using several particles with different surface hydrophobicities. In the measured force curves, a repulsive force was observed at large separation distances as the particle moved towards the bubble. The origin of the repulsive force was attributed to an electrostatic double-layer force because both the particle and bubble were negatively charged. After the repulsive force, an extremely long-range attractive force acted between the surfaces. These results indicate that the intervening thin water film between the particle and bubble rapidly collapsed, resulting in the particle penetrating the bubble.

The instability of the thin water film between the surfaces suggests the existence of an additional attractive force. By comparing the repulsive forces of the obtained force curves with the DLVO theory, the rupture thickness was estimated. The hydrophobicity of the particle did not significantly change the rupture thickness, whereas the pH of the solution is considered to be a critical factor.     

Direct measurement of the binding force between microfabricated particles and a planar surface in aqueous solution by force-sensing piezoresistive cantilevers

We propose a force measurement method for evaluating the binding force between microscale flat surfaces in an aqueous solution. Using force-sensing piezoresistive cantilevers with sub-nanonewton force resolution, we have directly measured binding forces between SiO2-SiO2 microcontacts, which were created by gravity-driven random collision between microfabricated SiO2 cylindrical particles and a planar SiO2 substrate in a HCl solution. First, to examine our method we measured the pH dependence of the binding force. The binding forces were 12 and 5.8 nN at pH 1.0 and 2.0, respectively. As the pH increased, the binding force decreased and became zero at pH greater than 3.0. We confirmed that the bindings were based on the van der Waals' (VDW) force at pH 2.0 or less whereas a repulsive double-layer force acted between the surfaces at pH 3.0 or more. Second, the binding forces were categorized into a friction force or an adhesion force between the particles and the substrate. In the measurement, the friction force between the particle and the substrate was measured in the case when the particle slid on the substrate. On the contrary, the adhesion force was measured when the particle came off the substrate. Whether the particle slid or came off depended on the aspect ratio of the particle. We fabricated cylindrical particles with an aspect ratio of 0.03-2.0 and distinguished the friction force from the adhesion force by changing the aspect ratio of the particles. As a result, the friction force per unit contact area between SiO2-SiO2 flat surfaces was found to be 330 pN/microm2 +/- 20% when we used particles with a low aspect ratio (<0.1), and the adhesion force per unit contact area was 90 pN/microm2 +/- 20% for particles with a high aspect ratio (>0.4). For fluidic self-assembly that utilizes microscale surface contact in a liquid, our measurement method is an effective tool for studying and developing systems.     

Rupture force between the third strand and the double strand within a triplex DNA

The rupture force to separate the third strand and the duplex within a triplex DNA was measured by means of atomic force spectroscopy. The tip and the sample surfaces were functionalized by oligodeoxyribonucleotides 5'-TTCTTCTTTCTTTTCCTTTTCTTTCTTCTTACTTCTCTCTCTC TCTCTCT-SH-3'. The sample surface was hybridized with 5'-AAGAAGAAAGAAAAGGAAAAGAAAGAAGAA-3' to form a double strand DNA on the surface prior to the force measurements. These sequences form triple helices with 30 base pairs under a pH of 5.8 and in the presence of 2.0 mM spermine. Signals of rupture of single and multiple triplex DNA were observed in the force distance curves. Rupture force histograms revealed a force of 42.6 +/- 1.9 pN from 24 independent measurements at a tip velocity of 400 nm/s to separate the third strand from duplex DNA. The velocity dependence of the rupture force quantum indicates a thermal dissociation process similar to that of rupturing a ds-DNA. The number of rupture events was controlled by adding oligonucleotides 5'-AAGAAGAAAGAAAAGGAAAAGAAAGAAGAA-3' either to reduce or to initiate triplex formation.     

Optimization of clamping force for low-viscosity polymer injection molding

Due to its low cost and high efficiency, injection molding is used for the mass production of many plastic products nowadays. However, when processing low-viscosity plastic materials, i.e., materials with an excellent fluidity, an inappropriate setting of the clamping force often results in a poor appearance and dimensional accuracy of the final product. Thus, operators usually take the upper limit of the clamping force as a default in setting up the machine in an attempt to improve the quality of the molded parts. However, such an approach shortens the machine and mold life, increases the energy consumption, and leads to poor air venting. Consequently, more scientific methods for determining the clamping force setting are required. To meet this demand, the present study proposes a clamping force search methodology for determining the optimal clamping force setting of a hydraulic cylinder clamping injection molding machine in the processing of low-viscosity plastics such as thermoplastic polyurethane (TPU) and polypropylene (PP). Based on the characteristic extracted from the sensing tie-bar elongation profile under different clamping force settings, a regression analysis on these data points is implemented to seek for an optimal clamping force. The experimental results show that for an injection molding machine with a hydraulic cylinder clamping mechanism, the effect of the mold temperature on the clamping force is sufficiently small to be ignored, which has an impact on the toggle type clamping unit. Furthermore, compared to traditional methods, the optimal clamping force obtained using the method proposed in the present study results in a significant improvement in the yield rate. Overall, the results confirm that for low-viscosity polymer resins, the optimal clamping force determined using the proposed method results in a higher and more consistent quality of the molded parts than that achieved using the proper clamping force setting for ordinary-viscosity resins.     

分子力场进展

分子力学（简称ＭＭ）是近年来化学家常用的一种计算方法。与量子力学从头计算和半经验方法相比，用分子力学处理大分子可以大大节省计算时间，而且，在大多数情况下，用分子力学方法计算得到的分子几何构型参数与实验值之间的差值可在实验误差范围之内。所以，分子力学是研究生物化学体系的有效和可行的手段。分子力学的核心是分子力场。本文介绍了分子力场的量子力学背景、分子力场和光谱力场之间的关系。分子力场的一般形式、分力     

Self-assembly of a dialkylurea gelator in organic solvents in the presence of centrifugal and shearing forces

A novel dialkylurea gelator, 1-methyl-2,4-bis(N(')-octadecaneureido)benzene (designated as MBOB) was synthesized, which can turn some organic solvents into organogels at extremely low concentrations (<2 wt%). The (1)H NMR spectra of MBOB in solution (110 degrees C) and in the gel state (30 degrees C) indicate that intermolecular hydrogen bonding is the driving force for the self-assembly of MBOB. In the process of the self-assembly of MBOB, orientation of MBOB aggregates occurs under the influence of external fields, such as a centrifugal force and shearing force fields. The minimum gelation concentrations of MBOB in organic solvents under a centrifugal force field were significantly higher than those in the absence of a centrifugal force field, indicating a significant effect of the external field on the self-assembly of MBOB. Field emission scanning electron microscopy (FE-SEM) provided evidence for a significantly phase transition of the MBOB aggregates from an amorphous state in the absence of the external field to an oriented state under conditions of a centrifugal or shearing force during the gelation process. A self-assembled structure of MBOB is proposed based upon an X-ray diffraction (XRD) analysis and a molecular simulation. DSC analysis of the organogels indicates that the phase transition temperature increased from 58.5 degrees C in the absence of the external field to 63.3 degrees C under a centrifugal force field and 62.2 degrees C under a shearing force field. The enthalpy of the phase transition decreased from 3.1 J/g in the absence of an external field to 2.6 J/g under a centrifugal force field and 2.7 J/g under a shearing force field.     

Hierarchical atom type definitions and extensible all‐atom force fields

The extensibility of force field is a key to solve the missing parameter problem commonly found in force field applications. The extensibility of conventional force fields is traditionally managed in the parameterization procedure, which becomes impractical as the coverage of the force field increases above a threshold. A hierarchical atom‐type definition (HAD) scheme is proposed to make extensible atom type definitions, which ensures that the force field developed based on the definitions are extensible. To demonstrate how HAD works and to prepare a foundation for future developments, two general force fields based on AMBER and DFF functional forms are parameterized for common organic molecules. The force field parameters are derived from the same set of quantum mechanical data and experimental liquid data using an automated parameterization tool, and validated by calculating molecular and liquid properties. The hydration free energies are calculated successfully by introducing a polarization scaling factor to the dispersion term between the solvent and solute molecules. © 2015 Wiley Periodicals, Inc.     

Chiral force constants: Recommendations for the presentation of internal coordinate force constants

Some force constants associated with the internal coordinates that sense handedness or chirality can have opposite signs in the enantiomers of chiral molecules. Examples of such force constants include interaction force constants between a torsional and stretching or bending internal coordinates. The sign reversal for these force constants in the enantiomers of chiral molecules or in opposite-handed molecular segments is best recognized by labeling them as chiral force constants. Recognition of chiral force constants suggests that certain guidelines are to be followed in the presentation of internal coordinate force constants. © 1993 by John Wiley & Sons, Inc. © John Wiley & Sons, Inc.     

Residence time, loading force, pH, and ionic strength affect adhesion forces between colloids and biopolymer-coated surfaces

Exopolymers are thought to influence bacterial adhesion to surfaces, but the time-dependent nature of molecular-scale interactions of biopolymers with a surface are poorly understood. In this study, the adhesion forces between two proteins and a polysaccharide [Bovine serum albumin (BSA), lysozyme, or dextran] and colloids (uncoated or BSA-coated carboxylated latex microspheres) were analyzed using colloid probe atomic force microscopy (AFM). Increasing the residence time of an uncoated or BSA-coated microsphere on a surface consistently increased the adhesion force measured during retraction of the colloid from the surface, demonstrating the important contribution of polymer rearrangement to increased adhesion force. Increasing the force applied on the colloid (loading force) also increased the adhesion force. For example, at a lower loading force of approximately 0.6 nN there was little adhesion (less than -0.47 nN) measured between a microsphere and the BSA surface for an exposure time up to 10 s. Increasing the loading force to 5.4 nN increased the adhesion force to -4.1 nN for an uncoated microsphere to a BSA surface and to as much as -7.5 nN for a BSA-coated microsphere to a BSA-coated glass surface for a residence time of 10 s. Adhesion forces between colloids and biopolymer surfaces decreased inversely with pH over a pH range of 4.5-10.6, suggesting that hydrogen bonding and a reduction of electrostatic repulsion were dominant mechanisms of adhesion in lower pH solutions. Larger adhesion forces were observed at low (1 mM) versus high ionic strength (100 mM), consistent with previous AFM findings. These results show the importance of polymers for colloid adhesion to surfaces by demonstrating that adhesion forces increase with applied force and detention time, and that changes in the adhesion forces reflect changes in solution chemistry.     

Effects of contact force and salt concentration on the unbinding of a DNA duplex by force spectroscopy

We report that varying the contact force in force spectroscopy results in a significant shift in DNA unbinding forces, measured from short oligonucleotides using a PicoForce microscope. The contact force between a 30-mer complementary DNA-coated probe and surface was varied from 100 pN to 10 nN, resulting in a significant shift in the most abundant unbinding force measured between the duplex. When contact forces were set at 200 pN or less, which is generally considered to be a low contact force region for biomolecular force spectroscopy studies, the shift in DNA unbinding forces was significant with changes in contact force. The effect of the salt concentration on the DNA unbinding forces was also examined for a range of salt concentrations from 5 to 500 mM because the presence of salt ions is necessary to facilitate the hybridization process. Although an increase in salt concentration resulted in the facilitation of DNA multiple binding events during force spectroscopy measurements, no significant shift in unbinding forces was observed. Our experiment demonstrates that the wide variation in DNA unbinding forces reported in the literature (50-600 pN) for short oligonucleotides can be accounted for by the different contact forces used and shows little or no effect of the salt concentration used in those studies. Furthermore, this study demonstrates the importance of reporting contact forces in force spectroscopy measurements for quantitative comparisons between different biomolecular systems, especially for noncovalent-type interactions.     

In situ hydrodynamic lateral force calibration of AFM colloidal probes

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.