Dual integrated actuators for extended range high speed atomic force microscopy

Cataloged from PDF version of article.A flexible system for increasing the throughput of the atomic force microscope without sacrificing imaging range is presented. The system is based on a nested feedback loop which controls a micromachined cantilever that contains both an integrated piezoelectric actuator and an integrated thermal actuator. This combination enables high speed imaging (2 mm/s) over an extended range by utilizing the piezoelectric actuator’s high bandwidth (15 kHz) and thermal actuator’s large response (300 nm/V). A constant force image, where the sample topography exceeds the range of the piezoelectric actuator alone, is presented. It has also been demonstrated that the deflection response of the thermal actuator can be linearized and controlled with an integrated diode. © 1999 American Institute of Physic

Amphotericin B induced interdigitation of apolipoprotein stabilized nanodisk bilayers

Amphotericin B nanodisks (AMB-ND) are ternary complexes of AMB, phospholipid (PL) and apolipoprotein organized as discrete nanometer scale disk-shaped bilayers. In gel filtration chromatography experiments, empty ND lacking AMB elute as a single population of particles with a molecular weight in the range of 200 kDa. AMB-ND formulated at a 4:1 PL:AMB weight ratio, separated into two peaks. Peak 1 eluted at the position of control ND lacking AMB while the second peak, containing all of the AMB present in the original sample, eluted in the void volume. When ND prepared with increased AMB (1:1 phospholipid:AMB molar ratio) were subjected to gel filtration chromatography, an increased proportion of phospholipid and apolipoprotein were recovered in the void volume with the AMB. Prior to gel filtration the AMB-ND sample could be passed through a 0.22 {micro}m filter without loss of AMB while the voided material was lost. Native gel electrophoresis studies corroborated the gel permeation chromatography data. Far UV circular dichroism analyses revealed that apoA-I associated with AMB-ND denatures at a lower guanidine HCl concentration than apoA-I associated with ND lacking AMB. Atomic force microscopy revealed that AMB induces compression of the ND bilayer thickness consistent with bilayer interdigitation, a phenomenon that is likely related to the ability of AMB to induce pore formation in susceptible membranes

A Compact, Low-Power Cantilever-Based Sensor Array for Chemical Detection

A compact and low-power cantilever-based sensor array has been developed and used to detect various vapor analytes. This device employs sorptive polymers that are deposited onto piezoresistive cantilevers. We have successfully detected several organic vapors, representing a breadth of chemical properties and over a range of concentrations. Comparisons of the polymer/vapor partition coefficient to the cantilever deflection responses show that a simple linear relationship does not exist, emphasizing the need to develop an appropriate functional model to describe the chemical-to-mechanical transduction that is unique to this sensing modality

Scan speed control for tapping mode SPM

In order to increase the imaging speed of a scanning probe microscope in tapping mode, we propose to use a dynamic controller on 'parachuting' regions. Furthermore, we propose to use variable scan speed on 'upward step' regions, with the speed determined by the error signal of the closed-loop control. We offer line traces obtained on a calibration grating with 25-nm step height, using both standard scanning and our scanning method, as experimental evidence

Parallel atomic force microscopy with optical interferometric detection

Cataloged from PDF version of article.We have developed an atomic force microscope that uses interferometry for parallel readout of a cantilever array. Each cantilever contains a phase sensitive diffraction grating consisting of a reference and movable set of interdigitated fingers. As a force is applied to the tip, the movable set is displaced and the intensity of the diffracted orders is altered. The order intensity from each cantilever is measured with a custom array of siliconphotodiodes with integrated complementary metal–oxide–semiconductor amplifiers. We present images from five cantilevers acquired in the constant height mode that reveal surface features 2 nm in height. The interdigital method for cantilever array readout is scalable, provides angstrom resolution, and is potentially simpler to implement than other methods. © 2001 American Institute of Physic

High-speed tapping mode imaging with active Q control for atomic force microscopy

Cataloged from PDF version of article.The speed of tapping mode imaging with the atomic force microscope(AFM) has been increased by over an order of magnitude. The enhanced operation is achieved by (1) increasing the instrument’s mechanical bandwidth and (2) actively controlling the cantilever’s dynamics. The instrument’s mechanical bandwidth is increased by an order of magnitude by replacing the piezotube z-axis actuator with an integrated zinc oxide (ZnO)piezoelectric cantilever. The cantilever’s dynamics are optimized for high-speed operation by actively damping the quality factor (Q) of the cantilever. Active damping allows the amplitude of the oscillating cantilever to respond to topography changes more quickly. With these two advancements, 80μm×80 μm high-speed tapping mode images have been obtained with a scan frequency of 15 Hz. This corresponds to a tip velocity of 2.4 mm/s. © 2000 American Institute of Physic

Dynamic force spectroscopy of parallel individual mucin1-antibody bonds

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

Single-Molecule Force Spectroscopy: Experiments, Analysis, and Simulations

International audienceThe mechanical properties of cells and of subcellular components are important to obtain a mechanistic molecular understanding of biological processes. The quantification of mechanical resistance of cells and biomolecules using biophysical methods matured thanks to the development of nanotechnologies such as optical and magnetic tweezers, the biomembrane force probe and atomic force microscopy (AFM). The quantitative nature of force spectroscopy measurements has converted AFM into a valuable tool in biophysics. Force spectroscopy allows the determination of the forces required to unfold protein domains and to disrupt individual receptor/ligand bonds. Molecular simulation as a computational microscope allows investigation of similar biological processes with an atomistic detail. In this chapter, we first provide a step-by-step protocol of force spectroscopy including sample preparation, measurement and analysis of force spectroscopy using AFM and its interpretation in terms of available theories. Next, we present the background for molecular dynamics (MD) simulations focusing on steered molecular dynamics (SMD) and the importance of bridging of computational tools with experimental technique