Michael Himmelhaus

Optical biosensor based on whispering gallery mode excitations in clusters of microparticles

A new concept for an optical biosensor based on whispering gallery mode (WGM) excitations in clusters of spherical microresonators is presented. Clusters of microresonators offer the advantage to exhibit specific WGM spectra that can be considered as their fingerprint. Therefore, individual clusters can be traced throughout an experiment even without knowledge of their precise positions. Polyelectrolyte adsorption onto clusters of 10μm polystyrene spheres is monitored in situ. It is shown that the WGMs shift to the same amount as those of a single microresonator and thus sensitivity does not depend on the number of microresonators present in the cluster.

Whispering gallery mode biosensor operated in the stimulated emission regime

Whispering gallery modes (WGMs) are generated in fluorescent polymer microparticles in phosphate buffered saline (PBS) above the threshold for stimulated emission and compared to their characteristics below threshold. The WGM microresonators show an eightfold improvement of their signal-to-noise ratio and a threefold increase in their quality factor when operated above threshold. In an investigation on the benefits for biochemical sensing, a real-time adsorption kinetics of bovine serum albumin in PBS is monitored and compared with those kinetics acquired by means of a WGM microresonator operated below threshold as well as by surface plasmon resonance (SPR).

In-vitro sensing of biomechanical forces in live cells by a whispering gallery mode biosensor

Direct measurement of the biomechanical stress induced by a live cell during endocytosis is reported. Fluorescent dye-doped polystyrene microspheres were used as microscopic remote optical sensors applying whispering gallery modes (WGMs) as transducer mechanism. Monitoring of the WGMs throughout the incorporation of the microsphere into the cell enabled the determination of the deformation experienced by the microsphere, characterized by both a broadening and a blue shift of the resonances, and consequently the stress induced by the cell. The results reveal an unexpectedly high stress with a magnitude of up to five times that of the passive cortical tension, which can be only explained by a so far undetermined active stress component induced by the cytoskeletal machinery during particle incorporation. The method is adaptable to the study of any other kind of phagocyte and thus provides a novel research tool of high interest for cell biology.

Interaction of self-assembled monolayers of oligo(ethylene glycol)-terminated alkanethiols with water studied by vibrational sum-frequency generation

Vibrational sum-frequency generation (VSFG) was used to investigate the conformational changes in self-assembled monolayers (SAMs) of (1-mercaptoundec-11-yl) hexa(ethylene glycol) monomethylether (EG6-OMe) on gold when exposed to liquid water. VSFG spectra of the EG6-OMe SAMs were recorded before, during, and after exposure of the films to water and after a subsequent evacuation step. While in contact with water the entire ethylene glycol chains are found in a random, solvated state, after removal from the fluid water molecules remain absorbed only at the terminal groups of the film giving rise to distinct conformational changes. After evacuation, the structure of the EG6-OMe SAM reverts to its original state, indicating that water has been removed from the monolayer. Our findings support recent ab initio calculations and Monte Carlo simulations on the interaction of ethylene glycol-terminated monolayers with water.

Optical Sensors Based on Whispering Gallery Modes in Fluorescent Microbeads: Response to Specific Interactions

Whispering gallery modes (WGMs) in surface-fixated fluorescent polystyrene microbeads are studied in view of their capability of sensing the formation of biochemical adsorption layers on their outer surface with the well-established biotin-streptavidin specific binding as the model system. Three different methods for analysis of the observed shifts in the WGM wavelength positions are applied and used to quantify the adsorbed mass densities, which are then compared with the results of a comparative surface plasmon resonance (SPR) study.

Optical Sensors Based on Whispering Gallery Modes in Fluorescent Microbeads: Size Dependence and Influence of Substrate

Whispering gallery modes in surface-fixated fluorescent polystyrene microbeads are studied in view of their capability of sensing changes in the refractive index of the beads’ environment by exposing them to water/glycerol mixtures of varying composition. The mode positions are analyzed by simultaneous fitting for mode number, bead radius, and environmental index. Down to a diameter of 8 μm, the sensor response follows the index of the bulk solution very well. For smaller bead sizes, some deviations occur, in particular for fluid indices not too different from that of water, which might be attributed to the presence of the substrate.

Label-free biosensors based on optically responsive nanocomposite layers: sensitivity and dynamic range.

Core-shell nanoparticle layers have proven to be a promising tool for the label-free detection of binding events. Upon reflection of white light, they exhibit pronounced extinction peaks in the UV/vis and NIR regime of the electromagnetic spectrum, which shift to higher wavelengths when molecules are adsorbed. Beside drastic simplification of the instrumentation and related reduction in cost, a significantly stronger response toward alkanethiol adsorption has been observed in previous experiments than in conventional surface plasmon resonance (SPR). However, as the amount of molecules deposited onto the nanoparticle films was unknown, no quantitative relationship could be established between the measured wavelength shifts and the surface mass density of the adsorbate. In order to facilitate quantitative molecule detection, self-assembled monolayers (SAMs) of simple and ethylene glycol (EG) terminated alkanethiols with various chain lengths were prepared on the nanoparticle-coated substrates. The measured red-shift of the extinction spectrum upon molecule adsorption was related to the amount of adsorbate as determined by X-ray photoelectron spectroscopy (XPS). For the whole range of film thicknesses studied, a linear relationship is found yielding a sensitivity factor of 0.027 nm/(ng/cm (2)). As proven by enzyme-linked immunosorbent assay (ELISA), such determined sensitivity factor can also be used to correctly predict the amount of surface-bound protein in immunoreactions from the measured wavelength shifts. It is concluded that the decay length of the evanescent electric field associated with the nanoparticle sensors is more than 100 nm and, thus, significantly larger than that observed for localized surface plasmons excited in small isolated metal clusters.

Confinement‐Induced Enhancement of Antigen–Antibody Interactions within Binary Nanopatterns to Achieve Higher Efficiency of On‐Chip Immunosensors

By combining molecular self-assembly, nanosphere lithography and reactive ion etching, large-scale nanopatterns of antibodies are fabricated for direct application in state-of-the-art on-chip immunosensors. Using in-situ surface plasmon resonance, the patterns are studied in view of their antigen binding capacity, which shows an increase of up to 120% solely in the case of antibody confinement into the nanopatches by means of a nonfouling embedding matrix.

Advances in label-free optical biosensing: direct comparison of whispering gallery mode sensors with surface plasmon resonance

Whispering gallery modes (WGM) in fluorescent dielectric microcavities have recently become an attractive alternative to state-of-the-art label-free optical biosensors due to their high sensitivity to molecular adsorption and their ease of operation under a variety of environmental conditions. In particular the true microscopic dimension of the sensor as well as its purely radiative control without any need for external coupling opens new opportunities for label-free biosensing on microscopic scale. While these are obvious advantages, a direct comparison of the performance of WGM biosensors with well-established techniques of known high sensitivity, such as surface plasmon resonance sensors, has not been undertaken to date, thus obscuring the opportunities of the newly rising approach. We have therefore studied the performance of both WGM biosensors and a commercial SPR sensor using a selection of specifically and non-specifically binding biomolecules in-situ and under same conditions. The WGM biosensors consist of 10 μm dye-doped polystyrene beads immobilized in a flow cell. The performance of the two techniques is compared in view of the efficiency and sensitivity towards detection of both model interaction pairs (e.g. biotin/Streptavidin) and specific interaction pairs such as antigen-antibody with a lower degree of interaction affinities.

Radial cavity mode excitation in fluorescent core-shell microcavities

In this letter the authors show that the optical mode spectrum of fluorescent dielectric microparticles can be entirely altered by coating the particles with a semitransparent gold layer. While the particles show the frequently observed whispering gallery mode (WGM) excitations without the metal shell, WGMs are suppressed after the coating procedure. Instead, the observed mode spectra can be explained by the evolution of radial modes, which the authors refer to as “spherical Fabry-Perot modes” in analogy to the mode patterns observed in planar Fabry-Perot interferometers.