Tangi Aubert

Hydrophilic, Bright CuInS2 Quantum Dots as Cd-Free Fluorescent Labels in Quantitative Immunoassay

We report on the synthesis of core-shell CuInS2/ZnS quantum dots (QDs) in organic solution, their encapsulation with a PEG-containing amphiphilic polymer, and the application of the resulting water-soluble QDs as fluorescent label in quantitative immunoassay. By optimizing the methods for core synthesis and shell growth, CuInS2/ZnS QDs were obtained with a quantum yield of 50% on average after hydrophilization. After conjugation with an aflatoxin B1-protein derivative, the obtained QDs were used as fluorescent labels in microplate immunoassay for the quantitative determination of the mycotoxin aflatoxin B1. QDs-based immunoassay showed higher sensitivity compared to enzyme-based immunoassay.

Cytotoxicity of cadmium-free quantum dots and their use in cell bioimaging.

The use of quantum dots (QDots) as bright and photostable probes for long-term fluorescence imaging is gaining more interest. Thus far, (pre)clinical use of QDots remains limited, which is primarily caused by the potential toxicity of QDots. Most QDots consist of Cd2+ ions, which are known to cause high levels of toxicity. In order to overcome this problem, several strategies have been tested, such as the generation of cadmium-free QDots. In the present study, two types of cadmium-free QDots, composed of ZnSe/ZnS (QDotZnSe) and InP/ZnS (QDotInP), were studied with respect to their cytotoxicity and cellular uptake in a variety of cell types. A multiparametric cytotoxicity approach is used, where the QDots are studied with respect to cell viability, oxidative stress, cell morphology, stem cell differentiation, and neurite outgrowth. The data reveal slight differences in uptake levels for both types of QDots (maximal for QDotZnSe), but clear differences in cytotoxicity and cell functionality effects exist, with highest toxicity for QDotZnSe. Differences between cell types and between both types of QDots can be explained by the intrinsic sensitivity of certain cell types and chemical composition of the QDots. At concentrations at which no toxic effects can be observed, the functionality of the QDots for fluorescence cell visualization is evaluated, revealing that the higher brightness of QDotZnSe overcomes most of the toxicity issues compared to that of QDotInP. Comparing the results obtained with common Cd2+-containing QDots tested under identical conditions, the importance of particle functionality is demonstrated, revealing that cadmium-free QDots tested in this study are not significantly better than Cd2+-containing QDots for long-term cell imaging and that more work needs to be performed in optimizing the brightness and surface chemistry of cadmium-free QDots for them to replace currently used Cd2+-containing QDots.

Synthesis and characterization of A4[Re6Q8L6]@SiO2 red-emitting silica nanoparticles based on Re6 metal atom clusters (A = Cs or K, Q = S or Se, and L = OH or CN).

Metal atom clusters constitute very promising candidates as luminophores for applications in biotechnology because they are nanosized entities offering robust luminescence in the near-infrared field (NIR). However, they cannot be used as prepared for biological applications because of potential toxic effects and quenching of the clusters luminescence in aqueous media, and they therefore need to be dispersed in a biocompatible matrix. We describe herein the encapsulation of octahedral rhenium clusters, denoted as A(4)[Re(6)Q(8)L(6)] (A = Cs or K, Q = S or Se, and L = OH or CN), in silica nanoparticles by a water-in-oil microemulsion process, paying particular attention to the clusters stability. The obtained A(4)[Re(6)Q(8)L(6)]@SiO(2) nanoparticles are 30 nm in size with good monodispersity and a perfectly spherical shape, as shown by scanning electron microscopy (SEM). The presence of cluster units inside the silica matrix was evidenced by scanning transmission electron microscopy in annular dark-field mode (ADF-STEM). From the point of view of their optical properties, the A(4)[Re(6)Q(8)L(6)]@SiO(2) nanoparticles show red and NIR emission under UV excitation, even when dispersed in water. The evolution of the structural and luminescence properties of clusters before and after encapsulation was followed by Raman and photoluminescence spectroscopy.

Silica-coated liposomes loaded with quantum dots as labels for multiplex fluorescent immunoassay

This manuscript describes synthesis and followed application of silica-coated liposomes loaded with quantum dots as a perspective label for immunoaasay. The hollow spherical structure of liposomes makes them an attractive package material for encapsulation of multiple water-insoluble quantum dots and amplifying the analytical signal. Silica coverage ensures the stability of the loaded liposomes against fusion and internal leakage during storage, transporting, application and also provides groups for bioconugation. For the first time these nanostructures were employed for the sensitive multiplex immunochemical determination of two analytes. As a model system mycotoxins zearalenone and aflatoxin B1 were detected in cereals. For simplification of multiassay results evaluation the silanized liposomed loaded with QDs of different colors were used. The IC50 values for the simultaneous determination of zearalenone and aflatoxin B1 were 16.2 and 18 µg kg(-1) for zearalenone and 2.2 and 2.6 µg kg(-1) for aflatoxin B1 in wheat and maize, respectively. As confirmatory method, liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was used.

Synthesis, modification, bioconjugation of silica coated fluorescent quantum dots and their application for mycotoxin detection.

To create bright and stable fluorescent biolabels for immunoassay detection of mycotoxin deoxynivalenol in food and feed, CdSe/CdS/ZnS core-shell quantum dots (QDs) were encapsulated in silica nanoparticles through a water-in-oil reverse microemulsion process. The optical properties and stability of the obtained silica coated QDs (QD@SiO2), modified with amino, carboxyl and epoxy groups and stabilized with polyethylene glycol fragments, were characterized in order to assess their bioapplicability. The developed co-condensation techniques allowed maintaining 80% of the initial fluorescent properties and yielded stable fluorescent labels that could be easily activated and bioconjugated. Further, the modified QD@SiO2 were efficiently conjugated with antibodies and applied as a novel label in a microtiter plate based immunoassay and a quantitative column-based rapid immunotest for deoxynivalenol detection with IC50 of 473 and 20 ng/ml, respectively.

Fast and versatile deposition of aligned semiconductor nanorods by dip-coating on a substrate with interdigitated electrodes

Semiconductor nanorods mainly absorb and emit light with the electric field along the axis of the rods, it is therefore important to align the nanorods along a preferred direction. The homogeneous deposition of aligned nanorods on large substrates is interesting for large size applications such as solar cells and OLEDs. In this work, we present a fast and versatile method for the homogeneous deposition and alignment of nanorods from a colloidal suspension. The method is based on a low-cost dip-coating procedure during which an alternating electric field is applied. The accumulation, orientation, and polarized fluorescence of the nanorods is verified by AFM and polarized fluorescence microscopy. An alignment with order parameter of 0.67 has been obtained with this method.

Nanoscale and Single-Dot Patterning of Colloidal Quantum Dots.

Using an optimized lift-off process we develop a technique for both nanoscale and single-dot patterning of colloidal quantum dot films, demonstrating feature sizes down to ~30 nm for uniform films and a yield of 40% for single-dot positioning, which is in good agreement with a newly developed theoretical model. While first of all presenting a unique tool for studying physics of single quantum dots, the process also provides a pathway toward practical quantum dot-based optoelectronic devices.

Low-loss silicon nitride waveguide hybridly integrated with colloidal quantum dots

Silicon nitride waveguides with a monolayer of colloidal quantum dots embedded inside were fabricated using a low-temperature deposition process and an optimized dry etching step for the composite layers. We experimentally demonstrated the luminescence of the embedded quantum dots is preserved and the loss of these hybrid waveguide wires is as low as 2.69dB/cm at 900nm wavelength. This hybrid integration of low loss silicon nitride photonics with active emitters offers opportunities for optical sources operating over a very broad wavelength range.

Active Liquid Crystal Tuning of Metallic Nanoantenna Enhanced Light Emission from Colloidal Quantum Dots

A system comprising an aluminum nanoantenna array on top of a luminescent colloidal quantum dot waveguide and covered by a thermotropic liquid crystal (LC) is introduced. By heating the LC above its critical temperature, we demonstrate that the concomitant refractive index change modifies the hybrid plasmonic-photonic resonances in the system. This enables active control of the spectrum and directionality of the narrow-band (∼6 nm) enhancement of quantum dot photoluminescence by the metallic nanoantennas.

Sensitive QD@SiO2-based immunoassay for triplex determination of cereal-borne mycotoxins

A sensitive tool for simultaneous quantitative determination of three analytes in one single well of a microtiter plate is shown for the first time. The developed technique is based on use of colloidal quantum dot enrobed into a silica shell (QD@SiO2) derivatives as a highly responsive label. Silica-coated quantum dots were prepared and subsequently modified via the co-hydrolysis with tetraethylorthosilicate (TEOS) and various organosilane reagents. Different surface modification schemes were compared in terms of applicability of the obtained particles for the multiplex immunoassay, e.g. stability and simplicity of their conjugation with biomolecules. As model system a multiplex immunosorbent assay for screening of three mycotoxins (deoxynivalenol, zearalenone and aflatoxin B1) in cereal-based products was realized via a co-immobilization of three different specific antibodies (anti- deoxynivalenol, anti-zearalenone and anti-aflatoxin B1) in one single well of a microtiter plate. Mycotoxins were simultaneously determined by labelling their conjugates with QD@SiO2 emitting in different parts of the visible spectrum. The limits of detection for the simultaneous determination were 6.1 and 5.3, 5.4 and 4.1, and 2.6 and 1.9µgkg(-1) for deoxynivalenol, zearalenone and aflatoxin B1 in maize and wheat, respectively. As confirmatory method, liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was used.