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Brahim Lounis

Publications and source records attributed to Brahim Lounis.

35 records · Page 2Linked to original sources

Quantum yield optimized fluorophores for site-specific labeling and super-resolution imaging

Single molecule applications, saturated pattern excitation microscopy, or stimulated emission depletion (STED) microscopy demand for bright and highly stable fluorescent dyes1,2. Despite of intensive research the choice of fluorphores is still very limited. Typically a stable fluorescent dyes is covalently attached to the target. This methodology brings forward a number of limitations, in particular, in case of protein labeling. First of all the fluorescent probes need to be attached selectively and site-specifically to prevent unspecific background. This often requires single cysteine mutations for covalent protein modification. Employing quantum dots allows overcoming problems of photo-bleaching3-6. However, the downsides are their large size, rendering the probe inaccessible to spatially confined architectures, issues in biocompatibility due to proper particle coating, and cellular toxicity6-8. Here we propose a new method to overcome the above outlined problems.

cond-mat.mes-hall↗

A New Route to Fluorescent SWNT/Silica Nanocomposites: Balancing Fluorescence Intensity and Environmental Sensitivity

We investigate the relationship between photoluminescence (PL) intensity and environmental sensitivity of surfactant-wrapped single walled carbon nanotubes (SWNTs). SWNTs were studied under a variety of conditions in suspension as well as encapsulated in silica nanocomposites, which were prepared by an efficient chemical vapor into liquids (CViL) sol-gel process. The dramatically improved silica encapsulation process described here has several advantages, including fast preparation and high SWNT loading concentration, over other encapsulation methods used to prepare fluorescent SWNT/silica nanocomposites. Further, addition of glycerol to SWNT suspensions prior to performing the CViL sol-gel process allows for the preparation of freestanding fluorescent silica xerogels, which to the best of our knowledge is the first report of such nanocomposites. Our spectroscopic data on SWNTs suspended in aqueous surfactants or encapsulated in silica show that achieving maximum PL intensity results in decreased sensitivity of SWNT emission response to changes imparted by the local environment. In addition, silica encapsulation can be used to "lock-in" a surfactant micelle structure surrounding SWNTs to minimize interactions between SWNTs and ions/small molecules. Ultimately, our work demonstrates that one should consider a balance between maximum PL intensity and the ability to sense environmental changes when designing new SWNT systems for future sensing applications.

cond-mat.mes-hall↗

All-optical trion generation in single walled carbon nanotubes

We present evidence of all optical trion generation and emission in undoped single walled carbon nanotubes (SWCNTs). Luminescence spectra, recorded on individual SWCNTs over a large CW excitation intensity range, show trion emission peaks red-shifted with respect to the bright exciton peak. Clear chirality dependence is observed for 22 separate SWCNT species, allowing for determination of electron-hole exchange interaction and trion binding energy contributions. Luminescence data together with ultrafast pump probe experiments on chirality sorted bulk samples suggest that exciton-exciton annihilation processes generate dissociated carriers that allow for trion creation upon a subsequent photon absorption event.

cond-mat.mes-hall↗

Efficient generation of near infra-red single photons from the zero-phonon line of a single molecule

Using the zero-phonon line (ZPL) emission of a single molecule, we realized a triggered source of near-infra-red (lambda=785 nm) single photons at a high repetition rate. A Weierstrass solid immersion lens is used to image single molecules with an optical resolution of 300 nm (~0.4*lambda) and a high collection efficiency. Because dephasing of the transition dipole due to phonons vanishes at liquid helium temperatures, our source is attractive for the efficient generation of single indistinguishable photons.

quant-ph↗

Indistinguishable near infra-red single photons from an individual organic molecule

By using the zero-phonon line emission of an individual organic molecule, we realized a source of indistinguishable single photons in the near infrared. A Hong-Ou-Mandel interference experiment is performed and a two-photon coalescence probability of higher than 50% at 2 K is obtained. The contribution of the temperature-dependent dephasing processes to the two-photon interference contrast is studied. We show that the molecule delivers nearly ideal indistinguishable single photons at the lowest temperatures when the dephasing is nearly lifetime limited. This source is used to generate post-selected polarization-entangled photon pairs, as a test-bench for applications in quantum information.

quant-ph↗

Luminescence Decay and the Absorption Cross-Section of Individual Single-Walled Carbon Nanotubes

The absorption cross section of highly luminescent individual single-walled carbon nanotubes is determined using time-resolved and cw luminescence spectroscopy. A mean value of 1x10-17 cm2 per carbon atom is obtained for (6,5) tubes excited at their second optical transition, and corroborated by single tube photothermal absorption measurements. Biexponential luminescence decays are systematically observed, with short and long lifetimes around 45 and 250 ps. This behavior is attributed to the band edge exciton fine structure with a dark level lying a few meV below a bright one.

cond-mat.mtrl-sci↗

Observation of intrinsic size effects in the optical response of individual gold nanoparticles

The Photothermal Heterodyne Imaging method is used to study for the first time the absorption spectra of individual gold nanoparticles with diameters down to 5 nm. Intrinsic size effects wich result in a broadening of the Surface Plasmon resonance are unambiguously observed. Dispersions in the peak energies and homogeneous widths of the single particle resonances are revealed. The experimental results are analysed within the frame of Mie theory.

physics.optics↗

Fluorescence microscopy of single autofluorescent proteins for cellular biology

In this paper we review the applicability of autofluorescent proteins for single-molecule imaging in biology. The photophysical characteristics of several mutants of the Green Fluorescent Protein (GFP) and those of DsRed are compared and critically discussed for their use in cellular biology. The alternative use of two-photon excitation at the single-molecule level or Fluorescence Correlation Spectroscopy is envisaged for the study of individual autofluorescent proteins. Single-molecule experiments performed in live cells using eGFP and preferably eYFP fusion proteins are reviewed. Finally, the first use at the single-molecule level of citrine, a more photostable variant of the eYFP is reported when fused to a receptor for neurotransmitter in live cells.

physics.optics↗

Multiple Routes for Glutamate Receptor Trafficking: Surface Diffusion and Membrane Traffic Cooperate to Bring Receptors to Synapses

Trafficking of glutamate receptors into and out of synapses is critically involved in the plasticity of excitatory synaptic transmission. Endocytosis and exocytosis of receptors have initially been thought to account alone for this trafficking. However, membrane proteins also traffic through surface lateral diffusion in the plasma membrane. We describe developments in electrophysiological and optical approaches that have allowed for the real time measurement of glutamate receptor surface trafficking in live neurons. These include (i) specific imaging of surface receptors using a pH sensitive fluorescent protein, (ii) design of a photoactivable drug to inactivate locally surface receptors and monitor electrophysiologically their recovery, and (iii)application of single molecule fluorescence microscopy to directly track the movement of individual surface receptors with nanometer resolution inside and outside synapses. Altogether, these approaches have demonstrated that glutamate receptors diffuse at high rates in the neuronal membrane and suggest a key role for surface diffusion in the regulation of receptor numbers at synapses.

physics.optics↗

Direct imaging of lateral movements of AMPA receptors inside synapses

Trafficking of AMPA receptors in and out of synapses is crucial for synaptic plasticity. Previous studies have focused on the role of endo/exocytosis processes or that of lateral diffusion of extra-synaptic receptors. We have now directly imaged AMPAR movements inside and outside synapses of live neurons using single-molecule fluorescence microscopy. Inside individual synapses, we found immobile and mobile receptors, which display restricted diffusion. Extra-synaptic receptors display free diffusion. Receptors could also exchange between these membrane compartments through lateral diffusion. Glutamate application increased both receptor mobility inside synapses and the fraction of mobile receptors present in a juxtasynaptic region. Block of inhibitory transmission to favor excitatory synaptic activity induced a transient increase in the fraction of mobile receptors and a decrease in the proportion of juxtasynaptic receptors. Altogether, our data show that rapid exchange of receptors between a synaptic and extra-synaptic localization occurs through regulation of receptor diffusion inside synapses.

physics.optics↗

Photothermal heterodyne imaging of individual nonfluorescent nanoclusters and nanocrystals

We introduce a new, highly sensitive, and simple heterodyne optical method for imaging individual nonfluorescent nanoclusters and nanocrystals. A 2 order of magnitude improvement of the signal is achieved compared to previous methods. This allows for the unprecedented detection of individual small absorptive objects such as metallic clusters (of 67 atoms) or nonluminescent semiconductor nanocrystals. The measured signals are in agreement with a calculation based on the scattering field theory from a photothermal-induced modulated index of refraction profile around the nanoparticle.

physics.optics↗

Photothermal Absorption Spectroscopy of Individual Semiconductor Nanocrystals

Photothermal heterodyne detection is used to record the first room-temperature absorption spectra of single CdSe/ZnS semiconductor nanocrystals. These spectra are recorded in the high cw excitation regime, and the observed bands are assigned to transitions involving biexciton and trion states. Comparison with the single nanocrystals photoluminescence spectra leads to the measurement of spectral Stokes shifts free from ensemble averaging.

physics.optics↗

Absorption spectroscopy of individual single-walled carbon nanotubes

Current methods for producing single-walled carbon nanotubes (SWNTs) lead to heterogeneous samples containing mixtures of metallic and semiconducting species with a variety of lengths and defects. Optical detection at the single nanotube level should thus offer the possibility to examine these heterogeneities provided that both SWNT species are equally well detected. Here, we used photothermal heterodyne detection to record absorption images and spectra of individual SWNTs. Because this photothermal method relies only on light absorption, it readily detects metallic nanotubes as well as the emissive semiconducting species. The first and second optical transitions in individual semicontucting nanotubes have been probed. Comparison between the emission and absorption spectra of the lowest-lying optical transition reveal mainly small Stokes shifts. Side bands in the near-infrared absorption spectra are observed and assigned to exciton-phonon bound states. No such sidebands are detected around the lowest transition of metallic nanotubes.

physics.optics↗

Single metallic nanoparticle imaging for protein detection in cells

We performed a visualization of membrane proteins labeled with 10-nm gold nanoparticles in cells, using an all-optical method based on photothermal interference contrast. The high sensitivity of the method and the stability of the signals allows 3D imaging of individual nanoparticles without the drawbacks of photobleaching and blinking inherent to fluorescent markers. A simple analytical model is derived to account for the measurements of the signal amplitude and the spatial resolution. The photothermal interference contrast method provides an efficient, reproducible, and promising way to visualize low amounts of proteins in cells by optical means.

physics.optics↗

Photothermal Heterodyne Imaging of Individual Metallic Nanoparticles: Theory versus Experiments

We present the theoretical and detailed experimental characterizations of Photothermal Heterodyne Imaging. An analytical expression of the photothermal heterodyne signal is derived using the theory of light scattering from a fluctuating medium. The amplitudes of the signals detected in the backward and forward configurations are compared and their frequency dependences are studied. The application of the Photothermal Heterodyne detection technique to the absorption spectroscopy of individual gold nanoparticles is discussed and the detection of small individual silver nanoparticles is demonstrated.

physics.optics↗

Single NanoParticle Photothermal Tracking (SNaPT) of 5 nm gold beads in live cells

Tracking individual nano-objets in live cells during arbitrary long times is an ubiquitous need in modern biology. We present here a method for tracking individual 5 nm gold nanoparticles on live cells. It relies on the photothermal effect and the detection of the Laser Induced Scattering around a NanoAbsorber (LISNA). The key point for recording trajectories at video rate is the use of a triangulation procedure. The effectiveness of the method is tested against Single fluorescent Molecule Tracking in live COS7 cells on subsecond time scales. We further demonstrate recordings for several minutes of AMPA receptors trajectories on the plasma membrane of live neurons. SNaPT has the unique potential to record arbitrary long trajectory of membrane proteins using non-fluorescent nanometer sized labels.

physics.bio-ph↗

Photothermal heterodyne imaging of individual non-fluorescent nano-objects

We introduce a new, highly sensitive, and simple heterodyne optical method for imaging individual non-fluorescent nano-objects. A two orders of magnitude improvement of the signal is achieved compared to previous methods. This allows for the unprecedented detection of individual small absorptive objects such as metallic clusters (of 67 atoms) or non-luminescent semiconductor nanocrystals. The measured signals are in agreement with a calculation based on the scattering field theory from a photothermal-induced modulated index of refraction profile around the nanoparticle.

cond-mat.other↗