Real-time continuous monitoring of dynamic concentration profiles studied with biosensing by particle motion.

Real-time monitoring-and-control of biological systems requires lab-on-a-chip sensors that are able to accurately measure concentration-time profiles with a well-defined time delay and accuracy using only small amounts of sampled fluid. Here, we study real-time continuous monitoring of dynamic concentration profiles in a microfluidic measurement chamber. Step functions and sinusoidal oscillations of concentrations were generated using two pumps and a herringbone mixer.

High-Throughput Single-Molecule Sensors: How Can the Signals Be Analyzed in Real Time for Achieving Real-Time Continuous Biosensing?

Single-molecule sensors collect statistics of single-molecule interactions, and the resulting data can be used to determine concentrations of analyte molecules. The assays are generally end-point assays and are not designed for continuous biosensing. For continuous biosensing, a single-molecule sensor needs to be reversible, and the signals should be analyzed in real time in order to continuously report output signals, with a well-controlled time delay and measurement precision.

Real-Time Detection of State Transitions in Stochastic Signals from Biological Systems.

Robust analysis of signals from stochastic biomolecular processes is critical for understanding the dynamics of biological systems. Measured signals typically show multiple states with heterogeneities and a wide range of state lifetimes. Here, we present an algorithm for robust detection of state transitions in experimental time traces where the properties of the underlying states are unknown.

Conformation switching of single native proteins revealed by nanomechanical probing without a pulling force.

Protein conformational changes are essential to biological function, and the heterogeneous nature of the corresponding protein states provokes an interest to measure conformational changes at the single molecule level. Here we demonstrate that conformational changes in single native proteins can be revealed by non-covalent antibody-targeting of specific domains within the protein, using nanomechanical probing without an applied pulling force. The protein of interest was captured between a particle and a substrate and three properties were quantified: the twist amplitude related to an applied torque, torsional compliance related to rotational Brownian motion, and translational Brownian displacement.

Continuous biomarker monitoring by particle mobility sensing with single molecule resolution.

Healthcare is in demand of technologies for real-time sensing in order to continuously guard the state of patients. Here we present biomarker-monitoring based on the sensing of particle mobility, a concept wherein particles are coupled to a substrate via a flexible molecular tether, with both the particles and substrate provided with affinity molecules for effectuating specific and reversible interactions. Single-molecular binding and unbinding events modulate the Brownian particle motion and the state changes are recorded using optical scattering microscopy.

Nanoscale Interparticle Distance within Dimers in Solution Measured by Light Scattering.

We demonstrate a novel approach to quantify the interparticle distance in colloidal dimers using Mie scattering. The interparticle distance is varied in a controlled way by changing the ionic strength of the solution and the magnetic attraction between the particles. The measured scaling behavior is interpreted using an energy-distance model that includes the repulsive electrostatic and attractive magnetic interactions.

Particle Motion Analysis Reveals Nanoscale Bond Characteristics and Enhances Dynamic Range for Biosensing.

Biofunctionalized colloidal particles are widely used as labels in bioanalytical assays, lab-on-chip devices, biophysical research, and in studies on live biological systems. With detection resolution going down to the level of single particles and single molecules, understanding the nature of the interaction of the particles with surfaces and substrates becomes of paramount importance. Here, we present a comprehensive study of motion patterns of colloidal particles maintained in close proximity to a substrate by short molecular tethers (40 nm).

Distance within colloidal dimers probed by rotation-induced oscillations of scattered light.

Aggregation processes of colloidal particles are of broad scientific and technological relevance. The earliest stage of aggregation, when dimers appear in an ensemble of single particles, is very important to characterize because it opens routes for further aggregation processes. Furthermore, it represents the most sensitive phase of diagnostic aggregation assays.

Probing the force-induced dissociation of aptamer-protein complexes.

Aptamers are emerging as powerful synthetic bioreceptors for fundamental research, diagnostics, and therapeutics. For further advances, it is important to gain a better understanding of how aptamers interact with their targets. In this work, we have used magnetic force-induced dissociation experiments to study the dissociation process of two different aptamer-protein complexes, namely for hIgE and Ara h 1.

High-efficiency dielectrophoretic ratchet.

Brownian ratchets enable the use of thermal motion in performing useful work. They typically employ spatial asymmetry to rectify nondirected external forces that drive the system out of equilibrium (cf. running marbles on a shaking washboard).

Quantification of protein-ligand dissociation kinetics in heterogeneous affinity assays.

Biochemical affinity assays inherently involve interactions of heterogeneous nature. We report a methodology to discriminate between and accurately characterize specific and nonspecific interactions in force-induced dissociation assays. Ligand-coupled superparamagnetic particles are incubated on surfaces coated with a mixture of specific receptors and nonspecifically interacting proteins.

One-step homogeneous magnetic nanoparticle immunoassay for biomarker detection directly in blood plasma.

Assay technologies capable of detecting low biomarker concentrations in complex biological samples are fundamental for biological research and for applications in medical diagnostics. In this paper we address the challenge to perform protein biomarker detection homogeneously in one single step, applying a minute amount of reagent directly into whole human blood plasma, avoiding any sample dilution, separation, amplification, or fluid manipulation steps. We describe a one-step homogeneous assay technology based on antibody-coated magnetic nanoparticles that are spiked in very small amount directly into blood plasma.

Roughness and ordering at the interface of oxidized polystyrene and water.

For the first time, atomistically detailed molecular dynamics calculations revealed molecular ordering of the water-oxidized atactic polystyrene (aPS) interface. Both ordering of the water molecules and the phenyl rings occur. In addition, the natural roughness of the surface has been simulated and compared to experimental values.

Frequency-selective rotation of two-particle nanoactuators for rapid and sensitive detection of biomolecules.

We describe an optomagnetic bionanotechnology for rapid and sensitive solution-based affinity assays. Nanoactuators made from bioactive magnetic nanoparticles undergo rotational motion in the volume of a fluid under frequency-controlled magnetic actuation. The nanoactuators show a time-dependent scattering cross-section to an incoming light beam.

Sensitivities of InGaAsP photonic crystal membrane nanocavities to hole refractive index.

The sensitivities of resonant wavelengths of photonic crystal (PhC) membrane nanocavities with embedded InAs quantum dots to the ambient refractive index are reported for use in (bio) chemical sensing. The resonances for the different modes of several point-defect type cavities are obtained by photoluminescence measurements. Systematic trends of the variation of sensitivity with increase of the overlap of the modes with the PhC holes are observed for varying cavity type as well as for a given mode within a cavity type.

Magnetically controlled rotation and torque of uniaxial microactuators for lab-on-a-chip applications.

We demonstrate the controlled rotation and torque generated by uniaxial magnetic microactuators formed by two bound superparamagnetic particles in a fluid. The torque and rotation are precisely controlled by rotating magnetic fields, generated by an external electromagnet or by on-chip current wires. We present the magnetic energy equations and the equations of motion for two-particle microactuators, with contributions from the permanent and induced magnetic moments of the particles.

Micro-fluidic actuation using magnetic artificial cilia.

We demonstrate advanced fluid manipulations using magnetic polymeric artificial cilia on the walls of a microfluidic channel. In nature, cilia are little hairs covering the surface of micro-organisms which enable them to manipulate a fluid on the micro-scale. The asymmetric movement of natural cilia is crucial to obtain a net fluid flow.

The mutual diffusion coefficient for (meth)acrylate monomers as determined with a nuclear microprobe.

The value of the mutual diffusion coefficient DV of two acrylic monomers is determined with nuclear microprobe measurements on a set of polymer films. These films have been prepared by allowing the monomers to diffuse into each other for a certain time and subsequently applying fast ultraviolet photo-polymerization, which freezes the concentration profile. The monomer diffusion profiles are studied with a scanning 2.