Quantifying Cell Binding Kinetics Mediated By Surface-Bound Blood Type B Antigen To Immobilized Antibodies

Cell adhesion is crucial to many biological processes, such as inflammatory responses, tumor metastasis and thrombosis formation. Recently a commercial surface plasmon resonance (SPR)-based BIAcore biosensor has been extended to determine cell binding mediated by surface-bound biomolecular interactions. How such cell binding is quantitatively governed by kinetic rates and regulating factors, however, has been poorly understood. Here we developed a novel assay to determine the binding kinetics of surface-bound biomolecular interactions using a commercial BIAcore 3000 biosensor. Human red blood cells (RBCs) presenting blood group B antigen and CM5 chip bearing immobilized anti-B monoclonal antibody (mAb) were used to obtain the time courses of response unit, or sensorgrams, when flowing RBCs over the chip surface. A cellular kinetic model was proposed to correlate the sensorgrams with kinetic rates. Impacts of regulating factors, such as cell concentration, flow duration and rate, antibody-presenting level, as well as pH value and osmotic pressure of suspending medium were tested systematically, which imparted the confidence that the approach can be applied to kinetic measurements of cell adhesion mediated by surface-bound biomolecular interactions. These results provided a new insight into quantifying cell binding using a commercial SPR-based BIAcore biosensor.

Low Spring Constant Regulates P-Selectin-Psgl-1 Bond Rupture

Forced dissociation of selectin-ligand bonds is crucial to such biological processes as leukocyte recruitment, thrombosis formation, and tumor metastasis. Although the bond rupture has been well known at high loading rate r(f) (>= 10(2) pN/s), defined as the product of spring constant k and retract velocity v, how the low r(f) (< 10(2) pN/s) or the low k regulates the bond dissociation remains unclear. Here an optical trap assay was used to quantify the bond rupture at r(f) <= 20 pN/s with low k (similar to 10(-3)-10(-2) pN/nm) when P-selectin and P-selectin glycoprotein ligand 1 (PSGL-1) were respectively coupled onto two glass microbeads. Our data indicated that the bond rupture force f retained the similar values when r(f) increased up to 20 pN/s. It was also found that f varied with different combinations of k and v even at the same r(f). The most probable force, f

Tensile Tests Of Micro Anchors Anodically Bonded Between Pyrex Glass And Aluminum Thin Film Coated On Silicon Wafer

Micro anchor is a kind of typical structures in micro/nano electromechanical systems (MEMS/NEMS), and it can be made by anodic bonding process, with thin films of metal or alloy as an intermediate layer. At the relative low temperature and voltage, specimens with actually sized micro anchor structures were anodically bonded using Pyrex 7740 glass and patterned crystalline silicon chips coated with aluminum thin film with a thickness comprised between 50 nm and 230 nm. To evaluate the bonding quality, tensile pulling tests have been finished with newly designed flexible fixtures for these specimens. The experimental results exhibit that the bonding tensile strength increases with the bonding temperature and voltage, but it decreases with the increase of the thickness of Al intermediate layer. This kind of thickness effect of the intermediate layer was not mentioned in the literature on anodic bonding. (C) 2008 Elsevier Ltd. All rights reserved.

Measurement Of Fracture Toughness And Interfacial Shear Strength Of Hard And Brittle Cr Coating On Ductile Steel Substrate

The fracture toughness and interfacial adhesion properties of a coating on its substrate are considered to be crucial intrinsic parameters determining performance and reliability of coating-substrate system. In this work, the fracture toughness and interfacial shear strength of a hard and brittle Cr coating on a normal medium carbon steel substrate were investigated by means of a tensile test. The normal medium carbon steel substrate electroplated with a hard and brittle Cr coating was quasi-statically stretched to induce an array of parallel cracks in the coating. An optical microscope was used to observe the cracking of the coating and the interfacial decohesion between the coating and the substrate during the loading. It was found that the cracking of the coating initiated at critical strain, and then the number of the cracks of the coating per unit axial distance increased with the increase in the tensile strain. At another critical strain, the number of the cracks of the coating became saturated, i.e. the number of cracks per unit axial distance became a constant after this critical strain. Based on the experiment result, the fracture toughness of the brittle coating can be determined using a mechanical model. Interestingly, even when the whole specimen fractured completely under an extreme strain of the substrate, the interfacial decohesion or buckling of the coating on its substrate was completely absent. The test result is different from that appeared in the literature though the identical test method and the brittle coating/ductile metal substrate system are taken. It was found that this difference can be attributed to an important mechanism that the Cr coating on the steel substrate has a good adhesion, and the ultimate interfacial shear strength between the Cr coating and the steel substrate has exceeded the maximum shear flow strength level of the steel substrate. This result also indicates that the maximum shear flow strength level of the ductile steel substrate can be only taken as a lower bound estimate on the ultimate shear strength of the interface. This estimation of the ultimate interfacial shear strength is consistent with the theoretical analysis and prediction presented in the literature.

Role Of Vertical Component Of Surface Tension Of The Droplet On The Elastic Deformation Of Pdms Membrane

Poly(dimethylsiloxane) (PDMS) has been widely used in lab-on-a-chip and micro- total analysis systems (mu-TAS), thus wetting and electrowetting behaviors of PDMS are of great importance in these devices. PDMS is a kind of soft polymer material, so the elastic deformation of PDMS membrane by a droplet cannot be neglected due to the vertical component of the interfacial tension between the liquid and vapor, and this vertical component of liquid-vapor surface tension is also balanced by the stress distribution within the PDMS membrane. Such elastic deformation and stress distribution not only affect the exact measurement of contact angle, but also have influence on the micro-fluidic behavior of the devices. Using ANSYS code, we simulated numerically the elastic deformation and stress distribution of PDMS membrane on a rigid substrate due to the liquid-vapor surface tension. It is found that the vertical elastic deformation of the PDMS membrane is on the order of several tens of nanometers due to the application of a droplet with a diameter of 2.31 mm, which is no longer negligible for lab-on-a-chip and mu-TAS. The vertical elastic deformation increases with the thickness of the PDMS membrane, and there exists a saturated membrane thickness, regarded as a semi-infinite membrane thickness, and the vertical elastic deformation reaches a limiting value when the membrane thickness is equal to or thicker than such saturated thickness. (C) Koninklijke Brill NV, Leiden, 2008.

Transitions Between Different Contact Models

The transitions between the different contact models which include the Hertz, Bradley, Johnson-Kendall-Roberts (JKR), Derjaguin-Muller-Toporov (DMT) and Maugis-Dugdale (MD) models are revealed by analyzing their contact pressure profiles and surface interactions. Inside the contact area, surface interaction/adhesion induces tensile contact pressure around the contact edge. Outside the contact area, whether or not to consider the surface interaction has a significant influence on the contact system equilibrium. The difference in contact pressure due to the surface interaction inside the contact area and the equilibrium influenced by the surface interaction outside the contact area are physically responsible for the different results of the different models. A systematic study on the transitions between different models is shown by analyzing the contact pressure profiles and the surface interactions both inside and outside the contact area. The definitions of contact radius and the flatness of contact surfaces are also discussed. (C) Koninklijke Brill NV, Leiden, 2008.

An Electrowetting Model For Rough Surfaces Under Low Voltage

Electrowetting is one of the most effective methods to enhance wettability. A significant change of contact angle for the liquid droplet can result from the surface microstructures and the external electric field, without altering the chemical composition of the system. During the electrowetting process on a rough surface, the droplet exhibits a sharp transition from the Cassie-Baxter to the Wenzel regime at a low critical voltage. In this paper, a theoretical model for electrowetting is put forth to describe the dynamic electrical control of the wetting behavior at the low voltage, considering the surface topography. The theoretical results are found to be in good agreement with the existing experimental results. (c) Koninklijke Brill NV, Leiden, 2008.

Interface Fracture Property Of Peo Ceramic Coatings On Aluminum Alloy

This paper combines the four-point bending test, SEM and finite element method to study the interface fracture property of PEO coatings on aluminum alloy. The interface failure mode of the coating on the compression side is revealed. The ceramic coating crack firstly along the 45 degrees to the interface, then the micro crack in the coating deduces the interface crack. The plastic deformation observed by SEM shows excellent adhesion property between the coating and substrate. The plastic deformation in the substrate is due to the interfacial crack extension, so the interface crack mode of PEO coatings is ductile crack. The results of FEM show that the compression strength is about 600 MPa. (C) 2008 Elsevier B.V. All rights reserved.

Extended Stoney'S Formula For A Film-Substrate Bilayer With The Effect Of Interfacial Slip

The curvature-stress relation is studied for a film-substrate bilayer with the effect of interfacial slip and compared with that of an ideal interface without interfacial slip. The interfacial slip together with the dimensions, elastic and interfacial properties of the film and substrate layers can cause a significant deviation of curvature-stress relation from that with an ideal interface. The interfacial slip also results in the so-called free edge effect that the stress, constraint force, and curvature vary dramatically around the free edges. The constant curvature as predicted by Stoney's formula and the Timoshenko model of an ideal interface is no longer valid for a bilayer with a nonideal interface. The models with the assumption of an ideal interface can also lead to an erroneous evaluation on the true stress state inside a bilayer with a nonideal interface. The extended Stoney's formula incorporating the effects of both the layer dimensions and interfacial slip is presented.

The Capillary Force In Micro- And Nano-Indentation With Different Indenter Shapes

The influence of the indenter shapes and various parameters on the magnitude of the capillary force is studied on the basis of models describing the wet adhesion of indenters and substrates joined by liquid bridges. In the former, we consider several shapes, such as conical, spherical and truncated conical one with a spherical end. In the latter, the effects of the contact angle, the radius of the wetting circle, the volume of the liquid bridge, the environmental humidity, the gap between the indenter and the substrate, the conical angle, the radius of the spherical indenter, the opening angle of the spherical end in the truncated conical indenter are included. The meniscus of the bridge is described using a circular approximation, which is reasonable under some conditions. Different dependences of the capillary force on the indenter shapes and the geometric parameters are observed. The results can be applicable to the micro- and nano-indentation experiments. It shows that the measured hardness is underestimated due to the effect of the capillary force. (c) 2008 Elsevier Ltd. All rights reserved.

Two-Dimensional Kinetics Of Beta(2)-Integrin And Icam-1 Bindings Between Neutrophils And Melanoma Cells In A Shear Flow

Cell adhesion, mediated by specific receptor-ligand interactions, plays an important role in biological processes such as tumor metastasis and inflammatory cascade. For example, interactions between beta(2)-integrin ( lymphocyte function-associated antigen-1 and/or Mac-1) on polymorphonuclear neutrophils (PMNs) and ICAM-1 on melanoma cells initiate the bindings of melanoma cells to PMNs within the tumor microenvironment in blood flow, which in turn activate PMN-melanoma cell aggregation in a near-wall region of the vascular endothelium, therefore enhancing subsequent extravasation of melanoma cells in the microcirculations. Kinetics of integrin-ligand bindings in a shear flow is the determinant of such a process, which has not been well understood. In the present study, interactions of PMNs with WM9 melanoma cells were investigated to quantify the kinetics of beta(2)-integrin and ICAM-1 bindings using a cone-plate viscometer that generates a linear shear flow combined with a two-color flow cytometry technique. Aggregation fractions exhibited a transition phase where it first increased before 60 s and then decreased with shear durations. Melanoma-PMN aggregation was also found to be inversely correlated with the shear rate. A previously developed probabilistic model was modified to predict the time dependence of aggregation fractions at different shear rates and medium viscosities. Kinetic parameters of beta(2)-integrin and ICAM-1 bindings were obtained by individual or global fittings, which were comparable to respectively published values. These findings provide new quantitative understanding of the biophysical basis of leukocyte-tumor cell interactions mediated by specific receptor-ligand interactions under shear flow conditions.

Robust Nanoadhesion Under Torque

In this article, optimization of shear adhesion strength between an elastic cylindrical fiber and a rigid substrate under torque is studied. We find that when the radius of the fiber is less than a critical value, the bonding-breaking along the contact interface occurs uniformly, rather than by mode III crack propagation. Comparison between adhesion models under torque and tension shows that nanometer scale of fibers may have evolved to achieve optimization of not only the normal adhesive strength but also the shear adhesive strength in tolerance of possible contact flaws.

Non-Slipping Jkr Model For Transversely Isotropic Materials

A generalized plane strain JKR model is established for non-slipping adhesive contact between an elastic transversely isotropic cylinder and a dissimilar elastic transversely isotropic half plane, in which a pulling force acts on the cylinder with the pulling direction at an angle inclined to the contact interface. Full-coupled solutions are obtained through the Griffith energy balance between elastic and surface energies. The analysis shows that, for a special case, i.e., the direction of pulling normal to the contact interface, the full-coupled solution can be approximated by a non-oscillatory one, in which the critical pull-off force, pull-off contact half-width and adhesion strength can be expressed explicitly. For the other cases, i.e., the direction of pulling inclined to the contact interface, tangential tractions have significant effects on the pull-off process, it should be described by an exact full-coupled solution. The elastic anisotropy leads to an orientation-dependent pull-off force and adhesion strength. This study could not only supply an exact solution to the generalized JKR model of transversely isotropic materials, but also suggest a reversible adhesion sensor designed by transversely isotropic materials, such as PZT or fiber-reinforced materials with parallel fibers. (c) 2007 Elsevier Ltd. All rights reserved.

Laser-induced wavy pattern formation in metal thin films

Laser-induced well-ordered and controllable wavy patterns are constructed in the deposited metal thin film. The micrometer-sized structure and orientation of the wavy patterns can be controlled via scanning a different size of rectangle laser spot on the films. Ordered patterns such as aligned, crossed, and whirled wave structures were designed over large areas. This patterning technique may find applications in both exploring the reliability and mechanical properties of thin films, and fabricating microfluidic devices. (C) 2004 American Institute of Physics.

Competitive adsorption of collagen and bovine serum albumin - effect of the surface wettability

The competitive adsorption of collagen and bovine serum albumin (BSA) on surfaces with varied wettability was investigated with imaging ellipsometry, and ellipsometry. Silane modified silicon surfaces were used as substrates. The results showed that surface wettability had an important effect on protein competitive adsorption. With the decrease of surface wettability, the adsorption of collagen from the mixture solution of collagen and BSA decreased, while the adsorption of BSA increased. (C) 2003 Elsevier B.V. All rights reserved.