Calibrating evanescent-wave penetration depths for biological TIRF microscopy

Roughly half of a cells proteins are located at or near the plasma membrane. In this restricted space the cell senses its environment, signals to its neighbors and ex-changes cargo through exo- and endocytotic mechanisms. Ligands bind to receptors, ions flow across channel pores, and transmitters and metabolites are transported against con-centration gradients. Receptors, ion channels, pumps and transporters are the molecular substrates of these biological processes and they constitute important targets for drug discovery. Total internal reflection fluorescence microscopy suppresses background from cell deeper layers and provides contrast for selectively imaging dynamic processes near the basal membrane of live-cells. The optical sectioning of total internal reflection fluorescence is based on the excitation confinement of the evanescent wave generated at the glass-cell interface. How deep the excitation light actually penetrates the sample is difficult to know, making the quantitative interpretation of total internal reflection fluorescence data problematic. Nevertheless, many applications like super-resolution microscopy, colocalization, fluorescence recovery after photobleaching, near-membrane fluorescence recovery after photobleaching, uncaging or photo-activation-switching, as well as single-particle tracking require the quantitative interpretation of evanescent-wave excited images. Here, we review existing techniques for characterizing evanescent fields and we provide a roadmap for comparing total internal reflection fluorescence data across images, experiments, and laboratories.Comment: 18 text pages, 7 figures and one supplemental figur

Molecular Plasmonics: strong coupling at the low molecular density limit

We study the strong coupling between the molecular excited state and the plasmonic modes of silver hole arrays with a resonant frequency very close to the asymptotic line of the plasmonic dispersion relation, at the nonlinear regime. We demonstrate that the strong coupling regime can be achieved between the two sub-systems at low molecular densities with negligible damping of the electromagnetic field. Our results are supported by rigorous numerical simulations showing that the strong coupling is observed when the molecular transition lies within the nonlinear regime of the dispersion relation rather than the linear regime.Comment: submitted to PCCP, 7 pages and 3 pages supporting informatio

Collective Plasmonic-Molecular Modes in the Strong Coupling Regime

We demonstrate strong coupling between molecular excited states and surface plasmon modes of a slit array in a thin metal film. The coupling manifests itself as an anti-crossing behavior of the two newly formed polaritons. As the coupling strength grows, a new mode emerges, which is attributed to long range molecular interactions mediated by the plasmonic field. The new, molecular-like mode repels the polariton states, and leads to an opening of energy gaps both below and above the asymptotic free molecule energy.Comment: 8 pages, 6 figures, submitted to PR

How do electronic carriers cross Si-bound alkyl monolayers?

Electron transport through Si-C bound alkyl chains, sandwiched between n-Si and Hg, is characterized by two distinct types of barriers, each dominating in a different voltage range. At low voltage, current depends strongly on temperature but not on molecular length, suggesting transport by thermionic emission over a barrier in the Si. At higher voltage, the current decreases exponentially with molecular length, suggesting tunneling through the molecules. The tunnel barrier is estimated, from transport and photoemission data, to be ~1.5 eV with a 0.25me effective mass.Comment: 13 pages, 3 figure