Silver nanostructure sensing platform for maximum-contrast fluorescence cell imaging.

We present herein a silver nanostructure-assisted sensing platform which consists of a combined structure of Ag nanowire (NW) and nanodot (ND) array. Highly enhanced fluorescence from fluorophore is attributed to a strongly coupled optical near-field interaction between proximately located Ag NW and NDs. We obtained enhanced fluorescence intensity with up to 140 folds, as contrasted from background intensity, reaching a theoretical maximum value.

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.

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.

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.

Analyte concentration at the tip of a nanopipette.

Concentration of molecules within the tips of nanopipettes when applying a DC voltage is herein investigated using finite-element simulations. The ion concentrations and fluxes due to diffusion, electro-migration, and electro-osmotic flow, and the electric potential are determined by the simultaneous solution of the Nernst-Planck, Poisson, and Navier-Stokes equations within the water solution containing sodium and chloride ions and negatively charged molecules. The electric potential within the pipette glass wall is at the same time determined by the Poisson equation together with appropriate boundary conditions and accounts for a field effect through the wall.

Reduction of photobleaching and photodamage in single molecule detection: observing single actin monomer in skeletal myofibrils.

Recent advances in detector technology make it possible to achieve single molecule detection (SMD) in a cell. SMD avoids complications associated with averaging signals from large assemblies and with diluting and disorganizing proteins. However, it requires that cells be illuminated with an intense laser beam, which causes photobleaching and cell damage.

Interference of surface plasmon resonances causes enhanced depolarized light scattering from metal nanoparticles.

We show that the strongly depolarized light scattering from noble metal particles is a result of interference of two surface plasmon resonances on the same particle. The maximum depolarization occurs between two resonances. Under favorable conditions the anisotropy of the scattering light can be much lower than what is possible for dielectric particles.

Focused optical beams obtained at planar structures by an imaginary shift in position.

A theory for focused optical beams at planar structures is described. It is an extension of a previous theory based on summation of plane waves. The focused beam is obtained by an imaginary shift in the position vector of the plane waves.

Red blood cells do not attenuate the SPCE fluorescence in surface assays.

We describe the positive effect of surface plasmon-coupled fluorescence emission (SPCE) on the detection of a signal from a surface immunoassay in highly absorbing or/and scattering samples. A model immunoassay using fluorescently labeled anti-rabbit antibodies that bind to rabbit immunoglobulin on a silver surface was performed, and the signal was detected in the presence of various highly absorbing and/or scattering solutions or suspensions, such as hemoglobin solution, plastic beads, and red blood cells. The results showed that a highly absorbing solution consisting of small molecules (dye, hemoglobin) attenuates the SPCE signal approximately 2-3-fold.

Surface plasmon-coupled emission and Fabry-Perot resonance in the sample layer: A theoretical approach.

A theoretical approach is used to investigate the coupling of surface plasmon-coupled emission to Fabry-Perot resonance in the sample layer. Quantities investigated are emission angles, polarization, power levels, and fluorescence lifetimes. The results are compared to experimental findings.

Propensity of a circadian clock to adjust to the 24h day-night light cycle and its sensitivity to molecular noise.

The circadian clock of Drosophila melanogaster and its tendency to adjust to the day-night light cycle is simulated by deterministic and stochastic methods. The robustness of the locking to the light-cycle with respect to molecular noise is studied. It is found that within the model studied, the molecular noise in the stochastic simulation erases the finer injection-locking structures, stronger injection signals are needed and the locking has the character of prolonged locked time intervals with cycle slips in between.

Directional two-photon induced surface plasmon-coupled emission.

We measured a directional surface plasmon-coupled emission (SPCE) induced by a two-photon absorption. A 60 nm thick layer of poly(vinyl alcohol) film doped with rhodamine 123 was deposited on a silvered (50 nm Ag) glass slide, which was attached to a hemicylindrical glass prism. The 820 nm excitation from a femtosecond Ti:Sapphire laser was used either in reverse Kretschmann or Kretschmann configuration.

Trapping single molecules by dielectrophoresis.

We have trapped single protein molecules of R-phycoerythrin in an aqueous solution by an alternating electric field. A radio frequency voltage is applied to sharp nanoelectrodes and hence produces a strong electric field gradient. The resulting dielectrophoretic forces attract freely diffusing protein molecules.

Theory and simulation of surface plasmon-coupled directional emission from fluorophores at planar structures.

A theoretical approach to surface plasmon-coupled emission (SPCE) from planar structures is developed. It is used for simulations. The results are compared to experimental findings.

Optical trapping of single fluorescent molecules at the detection spots of nanoprobes.

We propose a scheme of optical trapping of fluorescent molecules, based on the strongly enhanced optical field due to surface plasmon resonances at laser illuminated metal tips or particles. A semiclassical approach is compared to a quantum-mechanical one. Attractive as well as repulsive forces are possible depending on the wavelength of the optical field.