A relocated technique of atomic force microscopy (AFM) samples and its application in molecular biology

A kind of simple atomic force microscopy (AFM) relocated technique, which takes advantage of homemade sample locator system, is used for investigating repeatedly imaging of some specific species on the whole substrate (over 1 x 1 cm(2)) with resolution about 400 nm. As applications of this sample locator system, single extended DNA molecules and plasmid DNA network are shown in different AFM operational modes: tapping mode and contact mode with different tips after the substrates have been moved.

Quantifying the Effects of Contact Duration, Loading Rate, and Approach Velocity on P-Selectin-PSGL-1 Interactions Using AFM

Kinetics and its regulation by extrinsic physical factors govern selectin-ligand interactions that mediate tethering and rolling of circulating cells on the vessel wall under hemodynamic forces. While the force regulation of off-rate for dissociation of selectin-ligand bonds has been extensively studied, much less is known about how transport impacts the on-rate for association of these bonds and their stability. We used atomic force microscopy (AFM) to quantify how the contact duration, loading rate, and approach velocity affected kinetic rates and strength of bonds of P-selectin interacting with P-selectin glycoprotein ligand I (PSGL-1). We found a saturable relationship between the contact time and the rupture force, a biphasic relationship between the adhesion probability and the retraction velocity, a piece-wise linear relationship between the rupture force and the logarithm of the loading rate, and a threshold relationship between the approach velocity and the rupture force. These results provide new insights into how physical factors regulate receptor-ligand interactions.

Experimental Investigation of the Velocity Effect on Adhesion Forces with an Atomic Force Microscope Atomic Force Microscope

Capillary forces are significantly dominant in adhesive forces measured with an atomic force microscope (AFM) in ambient air, which are always thought to be dependent on water film thickness, relative humidity, and the free energy of water film. We study the nature of the pull-off force on a variety of surfaces as a function of tip velocity. It is found that the capillary forces are of relatively strong dependence on tip velocity. The present experiment is expected to provide a better understanding of the work mechanism of AFM in ambient air.

Competitive protein adsorption studied with atomic force microscopy and imaging ellipsometry

The adsorption and competitive adsorption of collagen and bovine serum albumin (BSA) were directly visualized and quantified using atomic force microscopy (AFM) and imaging ellipsometry. Chemically modified silicon surfaces were used as hydrophilic and hydrophobic substrates. The results showed that collagen and BSA in single component solution adsorbed onto a hydrophobic surface two times more than that onto a hydrophilic surface. The competitive adsorption between collagen and BSA showed that serum albumin preferentially adsorbed onto a hydrophobic surface, while collagen on a hydrophilic surface. In the binary solution of BSA (1 mg/ml BSA) and collagen (0.1 mg/ml), nearly 100% of the protein adsorbed onto the hydrophobic surface was BSA, but on the hydrophilic surface only about 6% was BSA. Surface affinity was the main factor controlling the competitive adsorption.

Studying Aggregate in Lysozyme Solution by Atomic Force Microscope

The aggregates in lysozyme solution with different NaCl concentration were investigated by Atomic Force Microscope (AFM). The AFM images show that there exist lysozyme monomers, n-mers and clusters in lysozyme solution when the conditions are not suitable for crystal growth. In favorable conditions for crystal growth, the lysozyme clusters disappear and almost only monomers exist in solution.

Atomic force microscopy observations of the topography of regenerated silk fibroin aggregations

How fibroin molecules fold themselves and further self-assemble into aggregations with specific structures when the solution concentration increases is the key to understanding the natural silk-forming process of the silkworm. A regenerated Bombyx mori silk fibroin solution was prepared, and serially diluted solutions were coated on aminated coverslips. Atomic force microscopy (AFM) observations of the topography of fibroin molecules revealed a transformation from rodlike aggregations 100-200 nm long to small globules 50 mn in diameter with decreasing concentrations. When the incubation duration increased, the aggregations of fibroin molecules showed a self-assembling process, which was measured with AFM. In particular, after the molecules were incubated for more than 20 min, rodlike micelles formed and were distributed evenly on the surface of the aminated slides. Flow chamber technology was used to study the effect of the shear loading on the topography of the fibroin molecular aggregations. After a shear loading was applied, larger rodlike particles formed at a higher incubation concentration in comparison with those at a lower concentration and were obviously oriented along the direction of fluid flow.

Competitive adsorption between bovine serum albumin and collagen observed by atomic force microscope

Atomic force microscopy (AFM) was used to study the competitive adsorption between bovine serum albumin (BSA) and type I collagen on hydrophilic and hydrophobic silicon wafers. BSA showed a grain shape and the type I collagen displayed fibril-like molecules with relatively homogeneous height and width, characterized with clear twisting (helical formation). These AFM images illustrated that quite a lot of type I collagen appeared in the adsorption layer on hydrophilic surface in a competitive adsorption state, but the adsorption of BSA was more preponderant than that of type I collagen on hydrophobic silicon wafer surface. The experiments showed that the influence of BSA on type I collagen adsorption on hydrophilic surface was less than that on hydrophobic surface.

Growth of liquid bridge in AFM

Capillary forces are dominant in adhesive forces measured with an atomic force microscope (AFM) in ambient air, which are thought to be dependent on water film thickness, relative humidity and the free energy of the water film. In this paper, besides these factors, we study the nature of the 'pull-off' force on a variety of atmospheres as a function of the contact time. It is found that capillary forces strongly depend on the contact time. In lower relative humidity atmosphere, the adhesion force is almost independent of the contact time. However, in higher relative humidity, the adhesion force increases with the contact time. Based on the experiment and a model that we present in this paper, the growth of the liquid bridge can be seen as undergoing two processes: one is water vapour condensation; the other is the motion of the thin liquid film that is absorbed on the substrate. The experiment and the growth model presented in this paper have direct relevance to the working mechanism of AFM in ambient air.

Nonlinear dynamics of atomic force microscopy with intermittent contact

When the atomic force microscopy (AFM) in tapping mode is in intermittent contact with a soft substrate, the contact time can be a significant portion of a cycle, resulting in invalidity of the impact oscillator model, where the contact time is assumed to be infinitely small. Furthermore, we demonstrate that the AFM intermittent contact with soft substrate can induce the motion of higher modes in the AFM dynamic response. Traditional ways of modeling AFM (one degree of freedom (DOF) system or single mode analysis) are shown to have serious mistakes when applied to this kind of problem. A more reasonable displacement criterion on contact is proposed, where the contact time is a function of the mechanical properties of AFM and substrate, driving frequencies/amplitude, initial conditions, etc. Multi-modal analysis is presented and mode coupling is also shown. (c) 2006 Published by Elsevier Ltd.

用AFM研究杜仲抗真菌蛋白的晶体生长

随着后基因组时代的到来以及蛋白质组学研究的深入开展,研究蛋白质晶体生长成为生物化学和结构生物学领域一个广泛关注的课题。通过使用原子力显微镜(Atomic Force Microscope,简称AFM)对杜仲抗真菌蛋白(eucommia antifungal protein,简称EAFP)的晶体在有母液存在下原位实时动态地进行了晶面生长观察。研究结果表明:不同过饱和度对EAFP晶体生长形貌的影响较大,较高的过饱和度下生长很快,生长台阶密度高,较高的过饱和度下主要进行各向异性二维台阶的发生、发展,较低的过饱和度下主要采用螺旋位错的生长方式,当过饱和度极低时生长缓慢,且晶体表面有很多小孔存在,晶面生长很不完整;还对不同过饱和度下晶体生长速率进行了定量的测量,也反映了过饱和度对EAFP晶体生长的影响;同时对在AFM观察过程中由探针的扫描速度和方向对表面形貌的影响进行了讨论。

A thin liquid film and its effects in an atomic force microscopy measurement

Recently, it has been observed that a liquid film spreading on a sample surface will significantly distort atomic force microscopy (AFM) measurements. In order to elaborate on the effect, we establish an equation governing the deformation of liquid film under its interaction with the AFM tip and substrate. A key issue is the critical liquid bump height y(0c) at which the liquid film jumps to contact the AFM tip. It is found that there are three distinct regimes in the variation of y(0c) with film thickness H, depending on Hamaker constants of tip, sample and liquid. Noticeably, there is a characteristic thickness H* physically defining what a thin film is; namely, once the film thickness H is the same order as H* , the effect of film thickness should be taken into account. The value of H* is dependent on Hamaker constants and liquid surface tension as well as tip radius.

Interaction between single molecules of Mac-1 and ICAM-1 in living cells: An atomic force microscopy study

The interaction between integrin macrophage differentiation antigen associated with complement three receptor function (Mac-1) and intercellular adhesion molecule-1 (ICAM-1), which is controlled tightly by the ligand-binding activity of Mac-1, is central to the regulation of neutrophil adhesion in host defense. Several "inside-out" signals and extracellular metal ions or antibodies have been found to activate Mac-1, resulting in an increased adhesiveness of Mac-1 to its ligands. However, the molecular basis for Mac-1 activation is not well understood yet. In this work, we have carried out a single-molecule study of Mac-1/ICAM-1 interaction force in living cells by atomic force microscopy (AFM). Our results showed that the binding probability and adhesion force of Mac-1 with ICAM-1 increased upon Mac-1 activation. Moreover, by comparing the dynamic force spectra of different Mac-1 mutants, we expected that Mac-1 activation is governed by the downward movement of its alpha 7 helix. (c) 2007 Elsevier Inc. All rights reserved.

Analysis of atomic force microscopic results of InAs islands formed by molecular beam epitaxy

Atomic force microscopy (AFM) measurements of nanometer-sized islands formed by 2 monolayers of InAs by molecular beam epitaxy have been carried out and the scan line of individual islands was extracted from raw AFM data for investigation. It is found that the base widths of nanometer-sized islands obtained by AFM are not reliable due to the finite size and shape of the contacting probe. A simple model is proposed to analyze the deviation of the measured value From the real value of the base width of InAs islands. (C) 1998 Elsevier Science B.V. All rights reserved.

Atomic force microscopic study of low temperature induced disassembly of RecA-dsDNA filaments

The assembly and disassembly of RecA-DNA nucleoprotein filaments on double-stranded DNA (dsDNA) or single-stranded DNA (ssDNA) are important steps for homologous recombination and DNA repair. The assembly and disassembly of the nucleoprotein filaments are sensitive to the reaction conditions. In this work, we investigated different morphologies of the formed nucleoprotein filaments at low temperature under different solution conditions by atomic force microscopy (AFM). We found that low temperature and long keeping time could induce the incomplete disassembly of the formed nucleoprotein filaments.

GIUSAXS and AFM Studies on Surface Reconstruction of Latex Thin Films during Thermal Treatment

The structural evolution of a single-layer latex film during annealing was studied via grazing incidence ultrasmall-angle X-ray scattering (GIUSAXS) and atomic force microscopy (AFM). The latex particles were composed of a low-T-g (-54 degrees C) core (n-butylacrylate, 30 wt %) and a high-T-g (41 degrees C) shell (t-butylacrylate, 70 wt %) and had an overall diameter of about 500 nm. GIUSAXS data indicate that the q(y) scan at q(z) = 0.27 nm(-1) (out-of-plane scan) contains information about both the structure factor and the form factor. The GIUSAXS data on latex films annealed at various temperatures ranging from room temperature to 140 degrees C indicate that the structure of the latex thin film beneath the surface changed significantly. The evolution of the out-of-plane scan plot reveals the surface reconstruction of the film. Furthermore, we also followed the time-dependent behavior of structural evolution when the latex film was annealed at a relatively low temperature (60 degrees C) where restructuring within the film can be followed that cannot be detected by AFM, which detects only surface morphology.