Guy Voirin

Biosensors Based on Porous Cellulose Nanocrystal–Poly(vinyl Alcohol) Scaffolds

Cellulose nanocrystals (CNCs), which offer a high aspect ratio, large specific surface area, and large number of reactive surface groups, are well suited for the facile immobilization of high density biological probes. We here report functional high surface area scaffolds based on cellulose nanocrystals (CNCs) and poly(vinyl alcohol) (PVA) and demonstrate that this platform is useful for fluorescence-based sensing schemes. Porous CNC/PVA nanocomposite films with a thickness of 25-70 nm were deposited on glass substrates by dip-coating with an aqueous mixture of the CNCs and PVA, and the porous nanostructure was fixated by heat treatment. In a subsequent step, a portion of the scaffolds hydroxyl surface groups was reacted with 2-(acryloxy)ethyl (3-isocyanato-4-methylphenyl)carbamate to permit the immobilization of thiolated fluorescein-substituted lysine, which was used as a first sensing motif, via nucleophile-based thiol-ene Michael addition. The resulting sensor films exhibit a nearly instantaneous and pronounced change of their fluorescence emission intensity in response to changes in pH. The approach was further extended to the detection of protease activity by immobilizing a Förster-type resonance energy transfer chromophore pair via a labile peptide sequence to the scaffold. This sensing scheme is based on the degradation of the protein linker in the presence of appropriate enzymes, which separate the chromophores and causes a turn-on of the originally quenched fluorescence. Using a standard benchtop spectrometer to monitor the increase in fluorescence intensity, trypsin was detected at a concentration of 250 μg/mL, i.e., in a concentration that is typical for abnormal proteolytic activity in wound fluids.

Single-pad scheme for integrated optical fluorescence sensing.

A new scheme for integrated optical fluorescence sensing is presented. The principle is based on a planar waveguide containing multiple sensing units, each consisting of a single-pad grating coupler structure. Single-pad means that all the following functions are incorporated in one single pad: laser light input, excitation of the labeled analyte molecules, efficient collection of the emitted fluorescent light into the waveguide, background suppression, and coupling the guided wave out to the detector. The results demonstrate a high efficiency of the fluorescence light excitation and collection, as well as a good suppression of the volume background.

Novel integrated optical sensor based on a grating coupler triplet

A novel approach for accomplishing robust integrated optical biosensors is presented. The principle is based on a symmetric grating coupler structure with the inherent feature of compensating disturbances originating from different kinds of mechanical instabilities. The complete sensor system has no moving parts and provides the high sensitivities typical for integrated optical sensors based on grating couplers. The feasibility of this approach is demonstrated by determining the refractive index of liquids as well as by measuring the specific binding of biomolecules (anti-chicken IgG) to immobilized chicken IgG on the sensor chip surface.

Physiologically responsive, mechanically adaptive polymer optical fibers for optogenetics

The capability to deliver light to specific locations within the brain using optogenetic tools has opened up new possibilities in the field of neural interfacing. In this context, optical fibers are commonly inserted into the brain to activate or mute neurons using photosensitive proteins. While chronic optogenetic stimulation studies are just beginning to emerge, knowledge gathered in connection with electrophysiological implants suggests that the mechanical mismatch of conventional optical fibers and the cortical tissue may be a significant contributor to neuroinflammatory response. Here, we present the design and fabrication of physiologically responsive, mechanically adaptive optical fibers made of poly(vinyl alcohol) (PVA) that may mitigate this problem. Produced by a one-step wet-spinning process, the fibers display a tensile storage modulus E of ∼7000u2009u2009MPa in the dry state at 25°C and can thus readily be inserted into cortical tissue. Exposure to water causes a drastic reduction of E to ∼35u2009u2009MPa on account of modest swelling with the water. The optical properties at 470 and 590 were comparable with losses of 0.7±0.04u2009u2009dB/cm at 470 nm and 0.6±0.1u2009u2009dB/cm at 590 nm in the dry state and 1.1±0.1u2009u2009dB/cm at 470 nm and 0.9±0.3u2009u2009dB/cm at 590 nm in the wet state. The dry end of a partially switched fiber with a length of 10 cm was coupled with a light-emitting diode with an output of 10.1 mW to deliver light with a power density of >500u2009u2009mW/cm2 from the wet end, which is more than sufficient to stimulate neurons in vivo. Thus, even without a low-refractive index cladding, the physiologically responsive, mechanically adaptive optical fibers presented here appear to be a very useful new tool for future optogenetic studies.

Au-labeled antibodies to enhance the sensitivity of a refractometric immunoassay: Detection of cocaine

An integrated platform for a very sensitive detection of cocaine based on a refractometric biosensor is demonstrated. The system uses a waveguide grating biosensor functionalized with a cocaine multivalent antigen-carrier protein conjugate. The immunoassay scheme consists of the competitive binding of cocaine-specific antibodies to the immobilized conjugates. A 1000-fold enhancement of the sensors sensitivity is achieved when using gold conjugated monoclonal antibodies instead of free antibodies. Together with the optimization of the assay conditions, the setup is designed for a quick identification of narcotics using automated sampling. The results show that the presence of cocaine in a liquid sample could be identified down to a concentration of 0.7 nM within one minute. This value can be reduced even further when longer binding time is allowed (0.2 nM after 15 min). Application of the system to detection of narcotics at airport security control points is discussed.

Smart Textiles with Biosensing Capabilities

Real-time, on-body measurement using minimally invasive biosensors opens up new perspectives for diagnosis and disease monitoring. Wearable sensors are placed in close contact with the body, performing analyses in accessible biological fluids (wound exudates, sweat). In this context, a network of biosensing optical fibers woven in textile enables the fabric to measure biological parameters in the surrounding medium. Optical fibers are attractive in view of their flexibility and easy integration for on-body monitoring. Biosensing fibers are obtained by modifying standard optical fibers with a sensitive layer specific to biomarkers. Detection is based on light absorption of the sensing fiber, placing a light source and a detector at both extremities of the fiber. Biosensing optical fibers have been developed for the in situ monitoring of wound healing, measuring pH and the activity of proteases in exudates. Other developments aim at the design of sensing patches based on functionalized, porous sol-gel layers, which can be deposited onto textiles and show optical changes in response to biomarkers. Biosensing textiles present interesting perspectives for innovative healthcare monitoring. Wearable sensors will provide access to new information from the body in real time, to support diagnosis and therapy.

Integrated optical biosensor for in-line monitoring of cell cultures.

An analytical detection platform was developed to evaluate the induced toxicity in cell cultures exposed to foreign agents like growth factors or nanoparticles. Connecting a biosensing detection device to the cell culture flasks allows analyzing the composition of cell medium in real-time. The analysis relies on the quantification of inflammatory cytokines released by cells into the cell culture medium, by means of solid-phase immunoassays analyzed with the wavelength interrogated optical sensing (WIOS) instrument. A fluidic system for in situ measurements allows detecting cytokines in real-time, with a sensitivity of 1-100 ng/mL depending on the cytokine. In addition, integration of an in-line optical absorbance measurement unit, in combination with the standard AB cell proliferation assay, provides information on the cell viability in the culture. Fluidic connections between the cell culture flasks, the optical biosensor and the absorbance measurement unit simultaneously allow quantifying up to three cytokines (interleukin 8, interleukin 6 and the monocyte chemotactic protein), assessing cellular proliferation, and thus discriminating between naïve cells and cells exposed to foreign agents such as growth factors (tumor necrosis factor alpha) or nanoparticles. This analytical tool presents a high potential for assessing the cytotoxicity of nanoparticles and other chemicals in vitro.

Design of nanostructured sol-gel coatings for (bio)sensing applications

Functionalized nanoporous coatings prepared by sol-gel processes are excellent sensing interfaces. Indeed, the high surface area combined with the specific functionalization using organic dyes and biologic entities has led to highly sensitive and selective optical (bio)sensors for gaseous or diluted species. Thus, biochips and optical fibers modified with functionalized nanoporous sensitive layers have been developed for environmental (e.g. gas detection, selective recognition of pesticides) and healthcare applications (e.g. wound monitoring).