903 resultados para Surface Enhanced Raman Spectroscopy


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This study compared the effect of physicochemical surface conditioning methods on the adhesion of bis-GMA-based resin cement to particulate filler composite (PFC) used for indirect dental restorations. PFC blocks (N (block)=54, n (block)=9 per group) were polymerized and randomly subjected to one of the following surface conditioning methods: a) No conditioning (Control-C), b) Hydrofluoric acid (HF)etching for 60s (AE60), c) HF for 90s (AE90), d) HF for 120s (AE120), e) HF for 180s (AE180), and f) air-abrasion with 30 mu m silica-coated alumina particles (AB). The conditioned surfaces were silanized with an MPS silane, and an adhesive resin was applied. Resin composite blocks were bonded to PFC using resin cement and photo-polymerized. PFC-cement-resin composite blocks were cut under coolant water to obtain bar specimens (1mmx0.8mm). Microtensile bond strength test (mu TBS)was performed in a universal testing machine (1mm/min). After debonding, failure modes were classified using stereomicroscopy. Surface characterization was performed on a set of separate specimen surfaces using Scanning Electron Microscopy (SEM), X-Ray Dispersive Spectroscopy (XDS), X-Ray Photoelectron Spectroscopy (XPS), and Fourier Transform-Raman Spectroscopy (FT-RS). Mean mu TBS (MPa) of C (35.6 +/- 4.9) was significantly lower than those of other groups (40.2 +/- 5.6-47.4 +/- 6.1) (p<0.05). The highest mu TBS was obtained in Group AB (47.4 +/- 6.1). Prolonged duration of HF etching increased the results (AE180: 41.9 +/- 7), but was not significantly different than that of AB (p>0.05). Failure types were predominantly cohesive in PFC (34 out of 54) followed by cohesive failure in the cement (16 out of 54). Degree of conversion (DC) of the PFC was 63 +/- 10%. SEM analysis showed increased irregularities on PFC surfaces with the increased etching time. Chemical surface analyses with XPS and FT-RS indicated 11-70% silane on the PFC surfaces that contributed to improved bond strength compared to Group C that presented 5% silane, which seemed to be a threshold. Group AB displayed 83% SiO2 and 17% silane on the surfaces.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Thin films of the bis[2,3,9,10,16,17,23,24-octachlorophthalocyaninate] lutetium(III) complex (LuPc2Cl32) have been prepared by the Langmuir-Blodgett and the Langmuir-Schaefer (LS) techniques. The influence of the chlorine substituents in the structure of the films and in their spectroscopic, electrochemical and sensing properties has been evaluated. The pi-A isotherms exhibit a monolayer stability greater than the observed in the unsubstituted analogue (LuPc2), being easily transferred to solid substrates, also in contrast to LuPc2. The LB and LS films present a linear growth forming stratified layers, monitored by UV-VIS absorption spectroscopy. The latter also revealed the presence of LuPc2Cl32 in the form of monomers and aggregates in both films. The FTIR data showed that the LuPc2Cl32 molecules present a non-preferential arrangement in both films. Monolayers of LB and LS were deposited onto 6 nm Ag island films to record surface-enhanced resonance Raman scattering (SERRS), leading to enhancement factors close to 2 x 10(3). Finally, LB and LS films deposited onto ITO glass have been successfully used as voltammetric sensors for the detection of catechol. The improved electroactivity of the LB and LS films has been confirmed by the reduction of the overpotential of the oxidation of catechol. The enhancement of the electrocatalytic effect observed in LB and LS films is the result of the nanostructured arrangement of the surface which increases the number of active sites. The sensors show a limit of detection in the range of 10(-5) mol/L.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Among the many methods developed for the synthesis of titanium dioxide, cathodic electrosynthesis has not received much attention because the resulting amorphous oxy-hydroxide matrix demands a further thermal annealing step to be transformed into crystalline titania. However, the possibility of filling deep recessed templates by the control of the solidliquid interface makes it a potentially suitable technique for the fabrication of porous scaffolds for photovoltaics and photocatalysis. Furthermore, a careful control of the crystallization process enables the growth of larger grains with lower density of grain boundaries, which act as electron traps that slow down electronic transport and promote charge recombination. In this report, well crystallized titania deposits were obtained by thermal annealing of amorphous deposits fabricated by cathodically assisted electrosynthesis on indium-tin oxide (ITO)substrates. The combined use of Raman spectroscopy and X-ray diffraction showed that the crystallization process is more intricate than previously assumed. It is shown that the amorphous matrix evolves into a rutile-free mixture of brookite and anatase at temperatures as low as 200 degrees C that persists up to 800 degrees C, when pure anatase dominates. The amount of brookite in the brookiteanatase mixture reaches a maximum at 400 degrees C. This very simple method for obtaining a brookiteanatase mixture and the ability to tune their proportions by thermal annealing is a promising alternative whose potential for solar cells and photocatalysis deserves a careful evaluation. Copyright (C) 2011 John Wiley & Sons, Ltd.

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This manuscript reports on the fabrication of plasmonic substrates using cathodic arc plasma ion implantation, in addition to their performance as SERS substrates. The technique allows for the incorporation of a wide layer of metallic nanoparticles into a polymer matrix, such as PMMA. The ability to pattern different structures using the PMMA matrix is one of the main advantages of the fabrication method. This opens up new possibilities for obtaining tailored substrates with enhanced performance for SERS and other surface-enhanced spectroscopies, as well as for exploring the basic physics of patterned metal nanostructures. The architecture of the SERS-active substrate was varied using three adsorption strategies for incorporating a laser dye (rhodamine): alongside the nanoparticles into the polymer matrix, during the polymer cure and within nanoholes lithographed on the polymer. As a proof-of-concept, we obtained the SERS spectra of rhodamine for the three types of substrates. The hypothesis of incorporation of rhodamine molecules into the polymer matrix during the cathodic arc plasma ion implantation was supported by FDTD (Finite-Difference Time-Domain) simulations. In the case of arrays of nanoholes, rhodamine molecules could be adsorbed directly on the gold surface, then yielding a well-resolved SERS spectrum for a small amount of analyte owing to the short-range interactions and the large longitudinal field component inside the nanoholes. The results shown here demonstrate that the approach based on ion implantation can be adapted to produce reproducible tailored substrates for SERS and other surface-enhanced spectroscopies.

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We describe a systematic investigation by the discrete dipole approximation on the optical properties of silver (Ag) and gold (Au) nanocubes as a function of the edge length in the 20-100 nm range. Our results showed that, as the nanocube size increased, the plasmon resonance modes shifted to higher wavelengths, the contribution from scattering to the extinction increased, and the quadrupole modes became more intense in the spectra. The electric field amplitudes at the surface of the nanocubes were calculated considering 514, 633 and 785 nm as the excitation wavelengths. While Ag nanocubes displayed the highest electric field amplitudes (vertical bar E vertical bar(max)) when excited at 514 nm, the Au nanocubes displayed higher vertical bar E vertical bar(max) values than Ag, for all sizes investigated, when the excitation wavelength was either 633 or 785 nm. The variations in vertical bar E vertical bar(max) as a function of size for both Ag and Au nanocubes could be explained based on the relative position of the surface plasmon resonance peak relative to the wavelength of the incoming electromagnetic wave. Our results show that not only size and composition, but also the excitation wavelength, can play an important role over the maximum near-field amplitudes values generated at the surface of the nanocubes.

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The development and characterization of biomolecule sensor formats based on the optical technique Surface Plasmon Resonance (SPR) Spectroscopy and electrochemical methods were investigated. The study can be divided into two parts of different scope. In the first part new novel detection schemes for labeled targets were developed on the basis of the investigations in Surface-plamon Field Enhanced Spectroscopy (SPFS). The first one is SPR fluorescence imaging formats, Surface-plamon Field Enhanced Fluorescence Microscopy (SPFM). Patterned self assembled monolayers (SAMs) were prepared and used to direct the spatial distribution of biomolecules immobilized on surfaces. Here the patterned monolayers would serve as molecular templates to secure different biomolecules to known locations on a surface. The binding processed of labeled target biomolecules from solution to sensor surface were visually and kinetically recorded by the fluorescence microscope, in which fluorescence was excited by the evanescent field of propagating plasmon surface polaritons. The second format which also originates from SPFS technique, Surface-plamon Field Enhanced Fluorescence Spectrometry (SPFSm), concerns the coupling of a fluorometry to normal SPR setup. A spectrograph mounted in place of photomultiplier or microscope can provide the information of fluorescence spectrum as well as fluorescence intensity. This study also firstly demonstrated the analytical combination of surface plasmon enhanced fluorescence detection with analyte tagged by semiconducting nano- crystals (QDs). Electrochemically addressable fabrication of DNA biosensor arrays in aqueous environment was also developed. An electrochemical method was introduced for the directed in-situ assembly of various specific oligonucleotide catcher probes onto different sensing elements of a multi-electrode array in the aqueous environment of a flow cell. Surface plasmon microscopy (SPM) is utilized for the on-line recording of the various functionalization steps. Hybridization reactions between targets from solution to the different surface-bound complementary probes are monitored by surface-plasmon field-enhanced fluorescence microscopy (SPFM) using targets that are either labeled with organic dyes or with semiconducting quantum dots for color-multiplexing. This study provides a new approach for the fabrication of (small) DNA arrays and the recording and quantitative evaluation of parallel hybridization reactions. In the second part of this work, the ideas of combining the SP optical and electrochemical characterization were extended to tethered bilayer lipid membrane (tBLM) format. Tethered bilayer lipid membranes provide a versatile model platform for the study of many membrane related processes. The thiolipids were firstly self-assembled on ultraflat gold substrates. Fusion of the monolayers with small unilamellar vesicles (SUVs) formed the distal layer and the membranes thus obtained have the sealing properties comparable to those of natural membranes. The fusion could be monitored optically by SPR as an increase in reflectivity (thickness) upon formation of the outer leaflet of the bilayer. With EIS, a drop in capacitance and a steady increase in resistance could be observed leading to a tightly sealing membrane with low leakage currents. The assembly of tBLMs and the subsequent incorporation of membrane proteins were investigated with respect to their potential use as a biosensing system. In the case of valinomycin the potassium transport mediated by the ion carrier could be shown by a decrease in resistance upon increasing potassium concentration. Potential mediation of membrane pores could be shown for the ion channel forming peptide alamethicin (Alm). It was shown that at high positive dc bias (cis negative) Alm channels stay at relatively low conductance levels and show higher permeability to potassium than to tetramethylammonium. The addition of inhibitor amiloride can partially block the Alm channels and results in increase of membrane resistance. tBLMs are robust and versatile model membrane architectures that can mimic certain properties of biological membranes. tBLMs with incorporated lipopolysaccharide (LPS) and lipid A mimicking bacteria membranes were used to probe the interactions of antibodies against LPS and to investigate the binding and incorporation of the small antimicrobial peptide V4. The influence of membrane composition and charge on the behavior of V4 was also probed. This study displays the possibility of using tBLM platform to record and valuate the efficiency or potency of numerous synthesized antimicrobial peptides as potential drug candidates.

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Membrane proteins play a major role in every living cell. They are the key factors in the cell’s metabolism and in other functions, for example in cell-cell interaction, signal transduction, and transport of ions and nutrients. Cytochrome c oxidase (CcO), as one of the membrane proteins of the respiratory chain, plays a significant role in the energy transformation of higher organisms. CcO is a multi centered heme protein, utilizing redox energy to actively transport protons across the mitochondrial membrane. One aim of this dissertation is to investigate single steps in the mechanism of the ion transfer process coupled to electron transfer, which are not fully understood. The protein-tethered bilayer lipid membrane is a general approach to immobilize membrane proteins in an oriented fashion on a planar electrode embedded in a biomimetic membrane. This system enables the combination of electrochemical techniques with surface enhanced resonance Raman (SERRS), surface enhanced reflection absorption infrared (SEIRAS), and surface plasmon spectroscopy to study protein mediated electron and ion transport processes. The orientation of the enzymes within the surface confined architecture can be controlled by specific site-mutations, i.e. the insertion of a poly-histidine tag to different subunits of the enzyme. CcO can, thus, be oriented uniformly with its natural electron pathway entry pointing either towards or away from the electrode surface. The first orientation allows an ultra-fast direct electron transfer(ET) into the protein, not provided by conventional systems, which can be leveraged to study intrinsic charge transfer processes. The second orientation permits to study the interaction with its natural electron donor cytochrome c. Electrochemical and SERR measurements show conclusively that the redox site structure and the activity of the surface confined enzyme are preserved. Therefore, this biomimetic system offers a unique platform to study the kinetics of the ET processes in order to clarify mechanistic properties of the enzyme. Highly sensitive and ultra fast electrochemical techniques allow the separation of ET steps between all four redox centres including the determination of ET rates. Furthermore, proton transfer coupled to ET could be directly measured and discriminated from other ion transfer processes, revealing novel mechanistic information of the proton transfer mechanism of cytochrome c oxidase. In order to study the kinetics of the ET inside the protein, including the catalytic center, time resolved SEIRAS and SERRS measurements were performed to gain more insight into the structural and coordination changes of the heme environment. The electrical behaviour of tethered membrane systems and membrane intrinsic proteins as well as related charge transfer processes were simulated by solving the respective sets of differential equations, utilizing a software package called SPICE. This helps to understand charge transfer processes across membranes and to develop models that can help to elucidate mechanisms of complex enzymatic processes.

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The two Mars Exploration Rovers (MER), Spirit and Opportunity, landed on the Martian surface in January 2004 and have since collected a wealth of information about their landing sites. As part of their payload, the miniaturised Mössbauer spectrometer MIMOS II contributes to the success of the mission by identifying Iron-bearing minerals and by determining Iron oxidation states in them. The basis of this work is the data set obtained at Opportunity’s landing site at Meridiani Planum. A portion of this data set is evaluated with different methods, with the aim to thoroughly characterize lithologic components at Meridiani Planum and possible relations between them.rnMIMOS II is able to measure Mössbauer spectra at different energies simultaneously, bearing information from different sampling depths of the investigated target. The ability of depth-selective Mössbauer spectroscopy to characterize weathered surface layers is illustrated through its application to two suitable rock targets that were investigated on Mars. In both cases, an enhanced concentration of Iron oxides at the rock surface was detected, pointing to a low degree of aqueous alteration. rnThe mineral hematite (α-Fe2O3) is present in the matrix of outcrop rocks and in spherules weathering from the outcrop. Simultaneous fitting of Mössbauer spectra was applied to data sets obtained on both target types to characterize the hematite component in detail. This approach reveals that two hematite populations are present, both in the outcrop matrix as well as in spherules. The hematite component with a comparably high degree of crystallinity and/or chemical purity is present in the outcrop matrix. The investigation of hematite at Meridiani Planum has shown that simultaneous fitting is a suitable and useful method to evaluate a large, correlated set of Mössbauer spectra.rnOpportunity encountered loose, cm-sized rocks along its traverse. Based on their composition and texture, these “cobbles” can be divided into three different groups. Outcrop fragments are impact-derived ejecta from local outcrop rocks. Cobbles of meteoritic origin contain the minerals kamacite (Fe,Ni) and troilite (FeS) and exhibit high Ni contents. Melt-bearing impact breccias bear similarities to local outcrop rocks and basaltic soil, with a phase composition and texture consistent with a formation scenario involving partial melting and inclusion of small, bright outcrop clasts. rnIron meteorites on the Martian surface experience weathering through the presence of even trace amounts of water due to their metallic nature. Opportunity encountered and investigated four Iron meteorites, which exhibit evidence for physical and chemical weathering. Discontinuous coatings contain Iron oxides, pointing to the influence of limited amounts of water. rnA terrestrial analogue site for Meridiani Planum is the Rio Tinto basin in south-west Spain. With its deposits of sulfate- and iron-oxide-bearing minerals, the region provides an adequate test bed for instrumentation for future Mars missions. In-situ investigations at Rio Tinto were carried out with a special focus on the combined use of Mössbauer spectroscopy with MIMOS II and Raman spectroscopy with a field-portable instrument. The results demonstrate that the two instruments provide complementary information about investigated samples.

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In this thesis, new advances in the development of spectroscopic based methods for the characterization of heritage materials have been achieved. As concern FTIR spectroscopy new approaches aimed at exploiting near and far IR region for the characterization of inorganic or organic materials have been tested. Paint cross-section have been analysed by FTIR spectroscopy in the NIR range and an “ad hoc” chemometric approach has been developed for the elaboration of hyperspectral maps. Moreover, a new method for the characterization of calcite based on the use of grinding curves has been set up both in MIR and in FAR region. Indeed, calcite is a material widely applied in cultural heritage, and this spectroscopic approach is an efficient and rapid tool to distinguish between different calcite samples. Different enhanced vibrational techniques for the characterisation of dyed fibres have been tested. First a SEIRA (Surface Enhanced Infra-Red Absorption) protocol has been optimised allowing the analysis of colorant micro-extracts thanks to the enhancement produced by the addition of gold nanoparticles. These preliminary studies permitted to identify a new enhanced FTIR method, named ATR/RAIRS, which allowed to reach lower detection limits. Regarding Raman microscopy, the research followed two lines, which have in common the aim of avoiding the use of colloidal solutions. AgI based supports obtained after deposition on a gold-coated glass slides have been developed and tested spotting colorant solutions. A SERS spectrum can be obtained thanks to the photoreduction, which the laser may induce on the silver salt. Moreover, these supports can be used for the TLC separation of a mixture of colorants and the analyses by means of both Raman/SERS and ATR-RAIRS can be successfully reached. Finally, a photoreduction method for the “on fiber” analysis of colorant without the need of any extraction have been optimised.

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Die Bildung kieselsäurehaltiger Spicula in marinen Schwämmen ist nur möglich durch die enzymatische Aktivität des Silicatein- in Verbindung mit der stöchiometrischen Selbstassemblierung des Enzyms mit anderen Schwammproteinen. Die vorliegende Arbeit basiert auf einem biomimetischen Ansatz mit dem Ziel, unterschiedliche Oberflächen für biotechnologische und biomedizinische Anwendungen mit Biosilica und Biotitania zu beschichten und zu funktionalisieren. Für biotechnologische Anwendungen ist dabei das Drucken von Cystein-getaggtem Silicatein auf Gold-Oberflächen von Bedeutung, denn es ermöglichte die Bildung definierter Biotitania-Strukturen (Anatas), welche als Photokatalysator den Abbau eines organischen Farbstoffs bewirkten. Des Weiteren zeigte sich die bio-inspirierte Modifikation von Tyrosin-Resten an rekombinantem Silicatein-(via Tyrosinase) als vielversprechendes Werkzeug zur Beschleunigung der Selbstassemblierung des Enzyms zu mesoskaligen Filamenten. Durch eine solche Modifikation konnte Silicatein auch auf der Oberfläche von anorganischen Partikeln immobilisiert werden, welches die Assemblierung von anorganisch-organischen Verbundwerkstoffen in wäßriger Umgebung förderte. Die resultierenden supramolekularen Strukturen könnten dabei in bio-inspirierten und biotechnologischen Anwendungen genutzt werden. Weiterhin wurde in der vorliegenden Arbeit die Sekundärstruktur von rekombinantem Silicatein- (Monomer und Oligomer) durch Raman Spektroskopie analysiert, nachdem das Protein gemäß einer neu etablierten Methode rückgefaltet worden war. Diese Spektraldaten zeigten insbesondere Änderungen der Proteinkonformation durch Solubilisierung und Oligomerisierung des Enzyms. Außerdem wurden die osteoinduzierenden und osteogenen Eigenschaften unterschiedlicher organischer Polymere, die herkömmlich als Knochenersatzmaterial genutzt werden, durch Oberflächenmodifikation mit Silicatein/Biosilica verbessert: Die bei der Kultivierung knochenbildender Zellen auf derart oberflächenbehandelten Materialien beobachtete verstärkte Biomineralisierung, Aktivierung der Alkalischen Phosphatase, und Ausbildung eines typischen zellulären Phänotyps verdeutlichen das Potential von Silicatein/Biosilica für der Herstellung neuartiger Implantat- und Knochenersatzmaterialien.

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L’électrofilage est une technique de mise en œuvre efficace et versatile qui permet la production de fibres continues d’un diamètre typique de quelques centaines de nanomètres à partir de l’application d’un haut voltage sur une solution concentrée de polymères enchevêtrés. L’évaporation extrêmement rapide du solvant et les forces d’élongation impliquées dans la formation de ces fibres leur confèrent des propriétés hors du commun et très intéressantes pour plusieurs types d’applications, mais dont on commence seulement à effleurer la surface. À cause de leur petite taille, ces matériaux ont longtemps été étudiés uniquement sous forme d’amas de milliers de fibres avec les techniques conventionnelles telles que la spectroscopie infrarouge ou la diffraction des rayons X. Nos connaissances de leur comportement proviennent donc toujours de la convolution des propriétés de l’amas de fibres et des caractéristiques spécifiques de chacune des fibres qui le compose. Les études récentes à l’échelle de la fibre individuelle ont mis en lumière des comportements inhabituels, particulièrement l’augmentation exponentielle du module avec la réduction du diamètre. L’orientation et, de manière plus générale, la structure moléculaire des fibres sont susceptibles d’être à l'origine de ces propriétés, mais d’une manière encore incomprise. L’établissement de relations structure/propriétés claires et l’identification des paramètres qui les influencent représentent des défis d’importance capitale en vue de tirer profit des caractéristiques très particulières des fibres électrofilées. Pour ce faire, il est nécessaire de développer des méthodes plus accessibles et permettant des analyses structurales rapides et approfondies sur une grande quantité de fibres individuelles présentant une large gamme de diamètre. Dans cette thèse, la spectroscopie Raman confocale est utilisée pour l’étude des caractéristiques structurales, telles que l’orientation moléculaire, la cristallinité et le désenchevêtrement, de fibres électrofilées individuelles. En premier lieu, une nouvelle méthodologie de quantification de l’orientation moléculaire par spectroscopie Raman est développée théoriquement dans le but de réduire la complexité expérimentale de la mesure, d’étendre la gamme de matériaux pour lesquels ces analyses sont possibles et d’éliminer les risques d’erreurs par rapport à la méthode conventionnelle. La validité et la portée de cette nouvelle méthode, appelée MPD, est ensuite démontrée expérimentalement. Par la suite, une méthodologie efficace permettant l’étude de caractéristiques structurales à l’échelle de la fibre individuelle par spectroscopie Raman est présentée en utilisant le poly(éthylène téréphtalate) comme système modèle. Les limites de la technique sont exposées et des stratégies expérimentales pour les contourner sont mises de l’avant. Les résultats révèlent une grande variabilité de l'orientation et de la conformation d'une fibre à l'autre, alors que le taux de cristallinité demeure systématiquement faible, démontrant l'importance et la pertinence des études statistiques de fibres individuelles. La présence de chaînes montrant un degré d’enchevêtrement plus faible dans les fibres électrofilées que dans la masse est ensuite démontrée expérimentalement pour la première fois par spectroscopie infrarouge sur des amas de fibres de polystyrène. Les conditions d'électrofilage favorisant ce phénomène structural, qui est soupçonné d’influencer grandement les propriétés des fibres, sont identifiées. Finalement, l’ensemble des méthodologies développées sont appliquées sur des fibres individuelles de polystyrène pour l’étude approfondie de l’orientation et du désenchevêtrement sur une large gamme de diamètres et pour une grande quantité de fibres. Cette dernière étude permet l’établissement de la première relation structure/propriétés de ces matériaux, à l’échelle individuelle, en montrant clairement le lien entre l’orientation moléculaire, le désenchevêtrement et le module d'élasticité des fibres.