Benoit Dubertret

Single-mismatch detection using gold-quenched fluorescent oligonucleotides

Here we describe a hybrid material composed of a single-stranded DNA (ssDNA) molecule, a 1.4 nm diameter gold nanoparticle, and a fluorophore that is highly quenched by the nanoparticle through a distance-dependent process. The fluorescence of this hybrid molecule increases by a factor of as much as several thousand as it binds to a complementary ssDNA. We show that this composite molecule is a different type of molecular beacon with a sensitivity enhanced up to 100-fold. In competitive hybridization assays, the ability to detect single mismatch is eightfold greater with this probe than with other molecular beacons.

Towards non-blinking colloidal quantum|[nbsp]|dots

At a single-molecule level, fluorophore emission intensity fluctuates between bright and dark states. These fluctuations, known as blinking, limit the use of fluorophores in single-molecule experiments. The dark-state duration shows a universal heavy-tailed power-law distribution characterized by the occurrence of long non-emissive periods. Here we have synthesized novel CdSe-CdS core-shell quantum dots with thick crystalline shells, 68% of which do not blink when observed individually at 33 Hz for 5 min. We have established a direct correlation between shell thickness and blinking occurrences. Importantly, the statistics of dark periods that appear at high acquisition rates (1 kHz) are not heavy tailed, in striking contrast with previous observations. Blinking statistics are thus not as universal as thought so far. We anticipate that our results will help to better understand the physico-chemistry of single-fluorophore emission and rationalize the design of other fluorophores that do not blink.

Colloidal nanoplatelets with two-dimensional electronic structure

The syntheses of strongly anisotropic nanocrystals with one dimension much smaller than the two others, such as nanoplatelets, are still greatly underdeveloped. Here, we demonstrate the formation of atomically flat quasi-two-dimensional colloidal CdSe, CdS and CdTe nanoplatelets with well-defined thicknesses ranging from 4 to 11 monolayers. These nanoplatelets have the electronic properties of two-dimensional quantum wells formed by molecular beam epitaxy, and their thickness-dependent absorption and emission spectra are described very well within an eight-band Pidgeon-Brown model. They present an extremely narrow emission spectrum with full-width at half-maximum less than 40 meV at room temperature. The radiative fluorescent lifetime measured in CdSe nanoplatelets decreases with temperature, reaching 1 ns at 6 K, two orders of magnitude less than for spherical CdSe nanoparticles. This makes the nanoplatelets the fastest colloidal fluorescent emitters and strongly suggests that they show a giant oscillator strength transition.

Cadmium-Free CuInS2/ZnS Quantum Dots for Sentinel Lymph Node Imaging with Reduced Toxicity

Semiconductor quantum dots (QDs) could significantly impact the performance of biomedical near-infrared (NIR) imaging by providing fluorescent probes that are brighter and more photostable than conventional organic dyes. However, the toxicity of the components of NIR emitting II-VI and IV-VI QDs that have been made so far (Cd, Hg, Te, Pb, etc.) has remained a major obstacle to the clinical use of QDs. Here, we present the synthesis of CuInS(2)/ZnS core/shell QDs emitting in the NIR ( approximately 800 nm) with good quantum yield and stability even after transfer into water. We demonstrate the potential of these QDs by imaging two regional lymph nodes (LNs) in vivo in mice. We then compare the inflammatory response of the axillary LN induced by different doses of CuInS(2)/ZnS and CdTeSe/CdZnS QDs and show a clear difference in acute local toxicity, the onset of inflammation only occurring at a 10 times more concentrated dose for CuInS(2)/ZnS QDs than for their Cd-containing counterparts.

Quasi 2D colloidal CdSe platelets with thicknesses controlled at the atomic level.

Well documented procedures to grow zero-dimensional systems, dots, and one-dimensional systems, wires and tubes, as colloidal particles in solution have been reported. In contrast, there are no methods of preparation that yield optically active two-dimensional soluble particles. Yet, ultrathin films (quantum wells) of II−VI and III−V semiconductors epitaxially grown on substrates by molecular beam epitaxy for example have proven extremely useful for both fundamental studies and a wealth of applications in optoelectronics. We show that II−VI cadmium selenide platelets, with thicknesses tuned at the atomic level, can be synthesized in solution. We describe the method for the preparation of these new colloidal nanocrystals and characterize them structurally and optically. We identified three platelets populations with emission maximums at 462, 513, and 550 nm with corresponding thicknesses estimated at 1.9, 2.2, and 2.5 nm, respectively. Despite the fact that the platelet aspect ratio within a population can...

Small and Stable Sulfobetaine Zwitterionic Quantum Dots for Functional Live-Cell Imaging

We have developed a novel surface coating for semiconductor quantum dots (QDs) based on a heterobifunctional ligand that overcomes most of the previous limits of these fluorescent probes in bioimaging applications. Here we show that QDs capped with bidentate zwitterionic dihydrolipoic acid-sulfobetaine (DHLA-SB) ligands are a favorable alternative to polyethylene glycol-coated nanoparticles since they combine small sizes, low nonspecific adsorption, preserved optical properties, and excellent stability over time and a wide range of pH and salinity. Additionally, these QDs can easily be functionalized with biomolecules such as streptavidin (SA) and biotin. We applied streptavidin-functionalized DHLA-SB QDs to track the intracellular recycling of cannabinoid receptor 1 (CB1R) in live cells. These QDs selectively recognized the pool of receptors at the cell surface via SA-biotin interactions with negligible nonspecific adsorption. The QDs retained their optical properties, allowing the internalization of CB1R into endosomes to be followed. Moreover, the cellular activity was apparently unaffected by the probe.

Core/Shell Colloidal Semiconductor Nanoplatelets

We have recently synthesized atomically flat semiconductor colloidal nanoplatelets with quasi 2D geometry. Here, we show that core/shell nanoplatelets can be obtained with a 2D geometry that is conserved. The epitaxial growth of the shell semiconductor is performed at room temperature. We report the detailed synthesis of CdSe/CdS and CdSe/CdZnS structures with different shell thicknesses. The shell growth is characterized both spectroscopically and structurally. In particular, the core/shell structure appears very clearly on high-resolution, high-angle annular dark-field transmission electron microscope images, thanks to the difference of atomic density between the core and the shell. When the nanoplatelets stand on their edge, we can precisely count the number of atomic planes forming the core and the shell. This provides a direct measurement, with atomic precision, of the core nanoplatelets thickness. The constraints exerted by the shell growth on the core is analyzed using global phase analysis. The core/shell nanoplatelets we obtained have narrow emission spectra with full-width at half-maximum close to 20 nm, and quantum yield that can reach 60%.

Synthesis, encapsulation, purification and coupling of single quantum dots in phospholipid micelles for their use in cellular and in vivo imaging

A detailed protocol for the synthesis of core/shell semiconductor nanocrystal, their encapsulation into phospholipid micelles, their purification and their coupling to a controlled number of small molecules is given. The protocol for the core/shell quantum dot (QD) CdSe/CdZnS synthesis has been specifically designed with two constraints in mind: green and reproducible core/shell QD synthesis with thick shell structure and QDs that can easily be encapsulated in poly(ethylene glycol)-phospholipid micelles with one QD per micelle. We present two procedures for the QD purification that are suitable for the use of QD micelles for in vivo imaging: ultracentrifugation and size-exclusion chromatography. We also discuss the different coupling chemistry for covalently linking a controlled number of molecules to the QD micelles. The total time durations for the different protocols are as follows: QD synthesis: 6 h; encapsulation: 15 min; purification: 1–4 h; coupling: reaction dependent.

Controlling spontaneous emission with plasmonic optical patch antennas.

We experimentally demonstrate the control of the spontaneous emission rate and the radiation pattern of colloidal quantum dots deterministically positioned in a plasmonic patch antenna. The antenna consists of a thin gold microdisk separated from a planar gold layer by a few tens of nanometers thick dielectric layer. The emitters are shown to radiate through the entire patch antenna in a highly directional and vertical radiation pattern. Strong acceleration of spontaneous emission is observed, depending on the antenna geometry. Considering the double dipole structure of the emitters, this corresponds to a Purcell factor up to 80 for dipoles perpendicular to the disk.

Design of new quantum dot materials for deep tissue infrared imaging

Near infrared fluorescence offers several advantages for tissue and in vivo imaging thanks to deeper photon penetration. In this article, we review a promising class of near infrared emitting probes based on semiconductor quantum dots (QDs), which have the potential to considerably improve in vivo fluorescence imaging thanks to their high brightness and stability. We discuss in particular the different criteria to optimize the design of near infrared QDs. We present the recent developments in the synthesis of novel QD materials and their different in vivo imaging applications, including lymph node localization, vasculature imaging, tumor localization, as well as cell tracking and QD-based multimodal probes.