A versatile dual spot laser scanning confocal microscopy system for advanced fluorescence correlation spectroscopy analysis in living cell

A fluorescence correlation spectroscopy (FCS) system based on two independent measurement volumes is presented. The optical setup and data acquisition hardware are detailed, as well as a complete protocol to control the location, size and shape of the measurement volumes. A method that allows to monitor independently the excitation and collection efficiency distribution is proposed. Finally, a few examples of measurements that exploit the two spots in static and/or scanning schemes, are reported.Comment: Accepted for publication in Review of Scientific Instrumen

Excitation Enhancement of a Quantum Dot Coupled to a Plasmonic Antenna

Plasmonic antennas are key elements to control the luminescence of quantum emitters. However, the antenna's influence is often hidden by quenching losses. Here, the luminescence of a quantum dot coupled to a gold dimer antenna is investigated. Detailed analysis of the multiply excited states quantifies the antenna's influence on the excitation intensity and the luminescence quantum yield separately

Field enhancement in a circular aperture surrounded by a single channel groove

International audienceNumerical analysis of diffraction by a single aperture surrounded by a circular shallow channel in a metallic screen shows the possibility of a 50-fold increase of the electric field intensity inside the central aperture, when compared to the incident field. Detailed analysis of cavity modes and their coupling through surface plasmon wave determine the parameters leading to maximum field enhancement. This effect can be used in high-efficiency single-molecule fluorescence analysis in attoliter volumes

Three-dimensional subwavelength confinement of light with dielectric microspheres

International audienceDielectric microspheres are shown to be capable of confining light in a three-dimensional region of subwavelength dimensions when they are illuminated by tightly focused Gaussian beams. We show that a simple configuration, not involving resonances, permits one to reach an effective volume as small as 0.6 (l/n)3. It is shown that this three-dimensional confinement arises from interferences between the field scattered by the sphere and the high angular components of the incident Gaussian beam passing aside the sphere

Increasing the lateral resolution of scanning microscopes by a factor of two using 2-Image microscopy

Increasing the resolution of optical microscopes is a challenging task for studying the cell machinery at the molecular level. 4Pi or TIRF microscopies permit one to reduce the axial dimension of the detection volume. To reduce its lateral dimension, we have proposed a solution in which the scanning head of a 4Pi microscope or of a confocal microscope is coupled to an interferometer. With this technique two beams coming from the source produce two images that are superimposed coherently. For this reason, one can call this technique 2-Image microscopy. It has been shown that with 2-Image microscopy, the complete use of the spatial frequencies collected by the objective allows to reach a 1.22 lambda/4NA lateral resolution. This improvement is independent of the excitation mode and is effective with incoherent light such as fluorescent or chemiluminescent (i.e. without optical excitation) samples. In this paper, we present an interferometric set-up and a modulation technique that make benefit fully from the advantages of 2-Image microscopy

Molecular fluorescence above metallic gratings

P. Andrew and William L. Barnes, Physical Review B, Vol. 64, article 125405 (2001). "Copyright © 2001 by the American Physical Society."We present measurements of the fluorescence of emitters located in close proximity (d<λ) to metallic grating surfaces. By measuring both the spontaneous emission lifetime and angle-dependent radiation pattern of a monolayer of dye molecules as a function of their separation from planar and periodically corrugated mirrors of increasing modulation depth, we are able to examine the effect of varying the surface profile on the emission process. Both the distance dependence of the lifetime and the spatial distribution of the emitted light are significantly changed upon the introduction of a corrugation, quite apart from the appearance of the familiar Bragg-scattered bound-mode features. It is postulated that these perturbations arise from the interference of the grating scattered dipole fields with the usual upward propagating and reflected fields. In addition, the measurement of nonexponential decay transients for the deepest gratings examined provide evidence for the existence of optically dissimilar dipole positions above the grating surface