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

Far-field radiation pattern in Coherent Anti-stokes Raman Scattering (CARS) Microscopy

Far field radiation pattern under tight focusing condition is investigated in Coherent Anti-stokes Raman Scattering (CARS) microscopy both in the forward (F-CARS) and backward (E-CARS) directions. While we assume no refraction index mismatch between the sample and the environing medium, our rigorous numerical electromagnetic computation takes into account the exact polarizations of the excitation laser beams and of the induced nonlinear dipoles. F-CARS and E-CARS radiation patterns, as well as their divergence, are studied as a function of the size of the sample object and compared to the excitation beams

Axial localization of luminophores by partial coherence interferometry

We propose a solution for increasing the axial resolution of confocal microscopes. In the experimental set-up described in this paper an interference phenomenon between two counterpropagating beams is used to determine the axial position of a luminophore. The optical path difference between the two waves, which is related to the position of the luminophore, is recovered thanks to a second interferometer by using partial coherence interferometry demodulation technique. The proposed solution can find applications in biology for localizing with nanometric resolution a small number of tagged species

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

Optical-fiber-microsphere for remote fluorescence correlation spectroscopy

International audienceFluorescence correlation spectroscopy (FCS) is a versatile method that would greatly benefit to remote optical-fiber fluorescence sensors. However, the current state-of-the-art struggles with high background and low detection sensitivities that prevent the extension of fiber-based FCS down to the single-molecule level. Here we report the use of an optical fiber combined with a latex microsphere to perform FCS analysis. The sensitivity of the technique is demonstrated at the single molecule level thanks to a photonic nanojet effect. This offers new opportunities for reducing the bulky microscope setup and extending FCS to remote or in vivo applications

Structure of molecular packing probed by polarization-resolved nonlinear four-wave mixing and coherent anti-Stokes Raman-scattering microscopy

International audienceWe report a method that is able to provide refined structural information on molecular packing in biomolecular assemblies using polarization-resolved four-wavemixing and coherent anti-Stokes Raman-scattering microscopy. These third-order nonlinear processes allow quantifying high orders of symmetry which are exploited here to reveal a high level of detail in the angular disorder behavior at the molecular scale in lipid membranes

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