Anne M. Charrier

Label free femtomolar electrical detection of Fe(III) ions with a pyridinone modified lipid monolayer as the active sensing layer

An innovative MOS-type field effect transistor was developed for the electrical detection of ferric ions. The sensing assays clearly show a specific detection with a gate-source voltage shift of up to 200 mV and a wide linear detection range (5 × 10-14 to 5 × 10-5 M) associated with good stability, selectivity and reproducibility.

Autonomic self-healing lipid monolayer: a new class of ultrathin dielectric.

The electrical performance of stabilized lipid monolayers on H-terminated silicon is reported for the first time. We show that these 2.7 nm thick only ultrathin layers present extremely low current leakage at high electric field and high breakdown voltage that both compare favorably with the best data reported on organic thin film dielectrics. We demonstrate a very unique property of autonomic self-healing of the layer at room temperature with the total recovery of its performance after electrical breakdown. The mechanisms involved in breakdown and self-healing are described.

Plasticity, elasticity, and adhesion energy of plant cell walls: nanometrology of lignin loss using atomic force microscopy

The complex organic polymer, lignin, abundant in plants, prevents the efficient extraction of sugars from the cell walls that is required for large scale biofuel production. Because lignin removal is crucial in overcoming this challenge, the question of how the nanoscale properties of the plant cell ultrastructure correlate with delignification processes is important. Here, we report how distinct molecular domains can be identified and how physical quantities of adhesion energy, elasticity, and plasticity undergo changes, and whether such quantitative observations can be used to characterize delignification. By chemically processing biomass, and employing nanometrology, the various stages of lignin removal are shown to be distinguished through the observed morphochemical and nanomechanical variations. Such spatially resolved correlations between chemistry and nanomechanics during deconstruction not only provide a better understanding of the cell wall architecture but also is vital for devising optimum chemical treatments.

Supported lipid monolayer with improved nanomechanical stability: effect of polymerization.

We study the effect of polymerization on the nanomechanical stability of supported lipid monolayers consisting of 1,2-di-(10Z,12Z-tricosadiynoyl)-sn-glycero-3-phosphocholine by means of force mapping using an atomic force microscope. For both nonpolymerized and polymerized lipid monolayers, we investigate the break-through forces required to rupture the monolayers for a whole range of loading velocities. We show that the average break-through force exerted by the tip and required to penetrate the monolayer has a logarithmic dependence on the loading rate. Both Young moduli and intrinsic Gibbs energies have been determined for the nonpolymerized and polymerized lipid monolayers, and we show a drastic effect of polymerization on the nanomechanical stability of the monolayer with an increase by a factor of ∼100 for the young modulus and ∼3 for the intrinsic Gibbs activation energy.

Size-Tunable Organic Nanodot Arrays: A Versatile Platform for Manipulating and Imaging Cells

Arrays of protein nanodots with dot-size tuned independently of spacing (e.g., ∼100 to 600 nm diameter for 900 nm spacing) are fabricated. The mechanism of size control is demonstrated, by numerical simulations, to arise from shadow effects during deposition of a sacrificial metal mask. We functionalize the nanodots with antibodies and embed them in a polymer-cushion or in lipid-bilayers or transfer them to soft elastomers. Their ability to influence cell architecture and local membrane organization is demonstrated in T-lymphocytes, using reflection interference contrast and total internal reflection fluorescence microscopy.

A field effect transistor biosensor with a γ-pyrone derivative engineered lipid-sensing layer for ultrasensitive Fe3+ ion detection with low pH interference.

Field effect transistors have risen as one of the most promising techniques in the development of biomedical diagnosis and monitoring. In such devices, the sensitivity and specificity of the sensor rely on the properties of the active sensing layer (gate dielectric and probe layer). We propose here a new type of transistor developed for the detection of Fe(3+) ions in which this sensing layer is made of a monolayer of lipids, engineered in such a way that it is not sensitive to pH in the acidic range, therefore making the device perfectly suitable for biomedical diagnosis. Probes are γ-pyrone derivatives that have been grafted to the lipid headgroups. Affinity constants derived for the chelator/Fe(3+) complexation as well as for other ions demonstrate very high sensitivity and specificity towards ferric ions with values as high as 5.10(10) M and a detected concentration as low as 50 fM.

Subpicomolar Iron Sensing Platform Based on Functional Lipid Monolayer Microarrays

We report herein the fabrication of novel microarrays based on air-stable functional lipid monolayers over silicon using a combination of e-beam lithography and lift-off. We demonstrate these microarrays can be use as ultrasensitive platform for Kelvin probe force microscopy in sensing experiments. Specificity of the detection is given by the functional group grafted at the lipid headgroup. The arrays developed for the detection of ferric ions, Fe(3+), using a γ-pyrone derivative chelator, demonstrate subpicomolar limit of detection with high specificity. In addition, the technique takes advantage of the structure of the array with the silicon areas playing the role of reference for the measurement, and we determine critical pattern dimensions below which the probe size/shape impacts the measured results.

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

Currently there is considerable interest in creating ordered arrays of adhesive protein islands in a sea of passivated surface for cell biological studies. In the past years, it has become increasingly clear that living cells respond, not only to the biochemical nature of the molecules presented to them but also to the way these molecules are presented. Creating protein micro-patterns is therefore now standard in many biology laboratories; nano-patterns are also more accessible. However, in the context of cell-cell interactions, there is a need to pattern not only proteins but also lipid bilayers. Such dual proteo-lipidic patterning has so far not been easily accessible. We offer a facile technique to create protein nano-dots supported on glass and propose a method to backfill the inter-dot space with a supported lipid bilayer (SLB). From photo-bleaching of tracer fluorescent lipids included in the SLB, we demonstrate that the bilayer exhibits considerable in-plane fluidity. Functionalizing the protein dots with fluorescent groups allows us to image them and to show that they are ordered in a regular hexagonal lattice. The typical dot size is about 800 nm and the spacing demonstrated here is 2 microns. These substrates are expected to serve as useful platforms for cell adhesion, migration and mechano-sensing studies.

Printing Functional Protein Nanodots on Soft Elastomers: From Transfer Mechanism to Cell Mechanosensing

Living cells sense the physical and chemical nature of their micro/nano environment with exquisite sensitivity. In this context, there is a growing need to functionalize soft materials with micro/nanoscale biochemical patterns for applications in mechanobiology. This, however, is still an engineering challenge. Here a new method is proposed, where submicronic protein-patterns are first formed on glass and are then printed on to an elastomer. The degree of transfer is shown to be governed mainly by hydrophobic interactions and to be influenced by grafting an appropriate fluorophore onto the core protein of interest. The transfer mechanism is probed by measuring the forces of adhesion/cohesion using atomic force microscopy. The transfer of functional arrays of dots with size down to about 400 nm, on elastomers with stiffness ranging from 3 kPa to 7 MPa, is demonstrated. Pilot studies on adhesion of T lymphocytes on such soft patterned substrates are reported.

Nanometrology of Biomass for Bioenergy: The Role of Atomic Force Microscopy and Spectroscopy in Plant Cell Characterization

Ethanol production using extracted cellulose from plant cell walls (PCW) is a very promising 4 approach to biofuel production. However, efficient throughput has been hindered by the pheno5 menon of recalcitrance, leading to high costs for the lignocellulosic conversion. To overcome 6 recalcitrance, it is necessary to understand the chemical and structural properties of the plant 7 biological materials, which have evolved to generate the strong and cohesive features observed 8 in plants. Therefore, tools and methods that allow the investigation of how the different molecular 9 components of PCW are organized and distributed and how this impacts the mechanical proper10 ties of the plants are needed but challenging due to the molecular and morphological complexity 11 of PCW. Atomic force microscopy (AFM), capitalizing on the interfacial nanomechanical forces, 12 encompasses a suite of measurement modalities for nondestructive material characterization. 13 Here, we present a review focused on the utilization of AFM for imaging and determination of 14 physical properties of plant-based specimens. The presented review encompasses the AFM 15 derived techniques for topography imaging (AM-AFM), mechanical properties (QFM), and sur16 face/subsurface (MSAFM, HPFM) chemical composition imaging. In particular, the motivation and 17 utility of force microscopy of plant cell walls from the early fundamental investigations to achieve 18 a better understanding of the cell wall architecture, to the recent studies for the sake of advancing 19 the biofuel research are discussed. An example of delignification protocol is described and the 20 changes in morphology, chemical composition and mechanical properties and their correlation at 21 the nanometer scale along the process are illustrated.