Nils Calander

Theory of surface-plasmon resonance optical-field enhancement at prolate spheroids

The optical-field enhancement from plasmon resonance at spheroids is studied by solving Maxwell equations using spheroidal vector wave functions. This treatment is an extension of the Mie theory for spheres. The phase retardation or dephasing effects, as studied by finite-element methods in a previous article, are confirmed. Nevertheless, the optical-field enhancement is shown to be substantial under certain resonance conditions. It is suggested that the positions of the resonances in parameter space are determined by global antenna properties and the magnitude of the field enhancement by local plasmon resonance.

Fluorescence correlation spectroscopy in surface plasmon coupled emission microscope

Study of dynamics of single molecules by Fluorescence Correlation Spectroscopy (FCS) requires that the rate of photon detection per molecule be high, that the background be low, and that there be a large change in fluorescent signal associated with change in a position of a molecule. FCS applied to microscopic Surface Plasmon Coupled Emission (SPCE) suggests a powerful method to meet those requirements. In this method, the observational volume is made shallow by placing a sample on a thin metal film and illuminating it with the laser beam at Surface Plasmon Resonance (SPR) angle through high numerical aperture objective. The illuminating light excites surface plasmons in the metal film that produce an evanescent wave on the aqueous side of the interface. The thickness of the detection volume is a product of evanescent wave penetration depth and distance-dependent fluorescence coupling to surface plasmons. It is further reduced by a metal quenching of excited fluorophores at a close proximity (below 10 nm) to a surface. The fluorescent light is emitted through the metal film only at an SPCE angle. Objective collects emitted light, and a confocal aperture inserted in its conjugate image plane reduces lateral dimensions of the detection volume to a fraction of a micrometer. By using diffusion of fluorescent microspheres, we show that SPCE-FCS is an efficient method to measure molecular diffusion and that on gold surface the height of the detection volume is ~35 nm.

Single molecule kinetics in the familial hypertrophic cardiomyopathy D166V mutant mouse heart

One of the sarcomeric mutations associated with a malignant phenotype of familial hypertrophic cardiomyopathy (FHC) is the D166V point mutation in the ventricular myosin regulatory light chain (RLC) encoded by the MYL2 gene. In this report we show that the rates of myosin cross-bridge attachment and dissociation are significantly different in isometrically contracting cardiac myofibrils from right ventricles of transgenic (Tg)-D166V and Tg-WT mice. We have derived the myosin cross-bridge kinetic rates by tracking the orientation of a fluorescently labeled single actin molecule. Orientation (measured by polarized fluorescence) oscillated between two states, corresponding to the actin-bound and actin-free states of the myosin cross-bridge. The rate of cross-bridge attachment during isometric contraction decreased from 3 s(-1) in myofibrils from Tg-WT to 1.4 s(-1) in myofibrils from Tg-D166V. The rate of detachment decreased from 1.3 s(-1) (Tg-WT) to 1.2 s(-1) (Tg-D166V). We also showed that the level of RLC phosphorylation was largely decreased in Tg-D166V myofibrils compared to Tg-WT. Our findings suggest that alterations in the myosin cross-bridge kinetics brought about by the D166V mutation in RLC might be responsible for the compromised function of the mutated hearts and lead to their inability to efficiently pump blood.

Plasmonic platforms of self-assembled silver nanostructures in application to fluorescence.

Fluorescence intensity changes were investigated theoretically and experimentally using self-assembled colloidal structures on silver semitransparent mirrors. Using a simplified quasi-static model and finite element method, we demonstrate that near-field interactions of metallic nanostructures with a continuous metallic surface create conditions that produce enormously enhanced surface plasmon resonances. The results were used to explain the observed enhancements and determine the optimal conditions for the experiment. The theoretical parts of the studies are supported with reports on detailed emission intensity changes which provided multiple fluorescence hot spots with 2-3 orders of enhancements. We study two kinds of the fluorophores: dye molecules and fluorescent nanospheres characterized with similar spectral emission regions. Using a lifetime-resolved fluorescence/reflection confocal microscopy technique, we find that the largest rate for enhancement (~1000-fold) comes from localized areas of silver nanostructures.

Monolayers of Silver Nanoparticles Decrease Photobleaching: Application to Muscle Myofibrils

Studying single molecules in a cell has the essential advantage that kinetic information is not averaged out. However, since fluorescence is faint, such studies require that the sample be illuminated with the intense light beam. This causes photodamage of labeled proteins and rapid photobleaching of the fluorophores. Here, we show that a substantial reduction of these types of photodamage can be achieved by imaging samples on coverslips coated with monolayers of silver nanoparticles. The mechanism responsible for this effect is the interaction of localized surface plasmon polaritons excited in the metallic nanoparticles with the transition dipoles of fluorophores of a sample. This leads to a significant enhancement of fluorescence and a decrease of fluorescence lifetime of a fluorophore. Enhancement of fluorescence leads to the reduction of photodamage, because the sample can be illuminated with a dim light, and decrease of fluorescence lifetime leads to reduction of photobleaching because the fluorophore spends less time in the excited state, where it is susceptible to oxygen attack. Fluorescence enhancement and reduction of photobleaching on rough metallic surfaces are usually accompanied by a loss of optical resolution due to refraction of light by particles. In the case of monolayers of silver nanoparticles, however, the surface is smooth and glossy. The fluorescence enhancement and the reduction of photobleaching are achieved without sacrificing the optical resolution of a microscope. Skeletal muscle myofibrils were used as an example, because they contain submicron structures conveniently used to define optical resolution. Small nanoparticles (diameter approximately 60 nm) did not cause loss of optical resolution, and they enhanced fluorescence approximately 500-fold and caused the appearance of a major picosecond component of lifetime decay. As a result, the sample photobleached approximately 20-fold more slowly than the sample on glass coverslips.

A subharmonic Josephson relaxation oscillator—amplification and locking

The inductance of the shunt loop in a resistively shunted Josephson tunnel junction may cause relaxation oscillations at subharmonics of the Josephson frequency. The relaxation period, the injection locking to a subharmonic of a strong external signal, and the amplification of weak signals were studied in detail. Gains up to 15 dB were achieved, but the noise temperature, which depended upon the harmonic of the relaxation frequency at which the amplification occurred, was high.

Shunted Josephson tunnel junctions: High‐frequency, self‐pumped low noise amplifiers

The high‐frequency amplification properties of transformer coupled, resistively shunted Josephson tunnel junctions have been investigated. The importance of the shunt loop inductance is stressed. It allows a high cutoff frequency, of significance for good high‐frequency performance. The self‐pumped parametric amplifier showed none of the excessive noise rise, which has hitherto plagued the development of externally pumped Josephson junction amplifiers. Around 10 GHz, we estimated a noise temperature less than 30 K for an amplifier pumped by a Josephson oscillation with a frequency well above twice the signal frequency. The corresponding gain of 5 dB may be increased in a better impedance matched circuit. The gain was very stable against variations in the bias conditions. A gain‐bandwidth product as high as 0.3 was registered. The experimental results agreed well with the established theory for self‐pumped parametric Josephson amplifiers. It should be possible to extend the low noise amplification by this ...

Molecular Detection and Analysis by Using Surface Plasmon Resonances

The surface plasmon resonance technique for molecular detection and analysis is reviewed, for example for monitoring binding of molecules to surfaces and for enhancing Raman spectroscopy. The possibility of modifying the emission and radiative decay rates of fluorophores by surface plasmon resonance is reviewed. Specifically, it concerns surface plasmon-coupled directional emission at planar structures.

dc characteristics of a nanoscale water-based transistor

We demonstrate a nanoscale water-based transistor. The presented nanoscale water-based transistor relies on the controlled modification of the pH in deionized water through the base applied electric field. The dc characteristics are presented and studied with a focus on the influence of the base applied electric field, the base electrode design, and their proximity to the sensing emitter and collector nanoelectrodes. The demonstrated water-based nanoscale device is of interest for many bioelectrical applications due to the biocompatibility and the wide usage and presence of water in biological systems.

Kinetics of a single cross-bridge in familial hypertrophic cardiomyopathy heart muscle measured by reverse Kretschmann fluorescence

Familial hypertrophic cardiomyopathy (FHC) is a serious heart disease that often leads to a sudden cardiac death of young athletes. It is believed that the alteration of the kinetics of interaction between actin and myosin causes FHC by making the heart to pump blood inefficiently. We set out to check this hypothesis ex vivo. During contraction of heart muscle, a myosin cross-bridge imparts periodic force impulses to actin. The impulses are analyzed by fluorescence correlation spectroscopy (FCS) of fluorescently labeled actin. To minimize observation volume and background fluorescence, we carry out FCS measurements in surface plasmon coupled emission mode in a reverse Kretschmann configuration. Fluorescence is a result of near-field coupling of fluorophores excited in the vicinity of the metal-coated surface of a coverslip with the surface plasmons propagating in the metal. Surface plasmons decouple on opposite sides of the metal film and emit in a directional manner as far-field p-polarized radiation. We show that the rate of changes of orientation is significantly faster in contracting cardiac myofibrils of transgenic mice than wild type. These results are consistent with the fact that mutated heart muscle myosin translates actin faster in in vitro motility assays.