Ecossistemas marinhos e estuarinos são o destino final de muitos contaminantes, e a ecotoxicologia ainda enfrenta a falta de organismos modelo para esses ambientes. Para desenvolver e apresentar um organismo teste adequado para ser utilizado em estudos de laboratório e campo, métodos para o cultivo, endpoints e biomarcadores devem ser avaliados. O anfípode marinho Parhyale hawaiensis é um modelo para estudos de biologia evolutiva e embrionária, é mundialmente distribuído e fácil de manipular em laboratório e consequentemente um modelo interessante para estudos ecotoxicológicos. O objetivo desse trabalho é padronizar condições de cultivo, teste de toxicidade, e desenvolver biomarcadores no anfípode P. hawaiensis e aplicá-los para avaliar os efeitos de nanopartículas metálicas na exposição via alimentação. As condições de cultivo foram estabelecidas em água salina reconstituída (salinidade 30 ± 2), temperatura de 24 ± 2 oC, fotoperíodo de 12-12h luz/escuro, coral triturado como substrato, alimentação diária com ração de peixe, troca de água parcial e aeração constante. As condições de teste compreendem a utilização de microplaca de 96-poços, organismos <7 dias, um organismo por poço com 200 &#181;L de solução por 96 h. Zinco, cobre, prata, cádmio e amônia foram selecionados como substância teste e a sensibilidade de P. hawaiensis foi similar e dentro da faixa de outros anfípodes marinhos. As condições para expressão gênica em P. hawaiensis foram desenvolvidas para o gene de referência 18S, dois genes de metalotioneínas (MT) e dois genes de glutationa S-transferase (GST). Entretanto, os genes selecionados não foram diferencialmente expressos nas condições testadas. Sendo assim, a análise da expressão gênica global, RNA-seq, foi realizada com organismos adultos alimentados por 7 dias com alimento contaminado com nanopartículas de prata (Ag-NPs), AgCl e controle. A análise da expressão gênica global (RNA-seq) foi conduzida com sucesso e destacou diferenças na resposta entre machos e fêmeas, bem como nos tratamentos com AgCl e Ag-NP. A análise comparativa revelou genes comuns entre as duas formas de Ag estudadas, e mostrou respostas diferentes no tratamento com Ag-NP, portanto, efeitos adversos diferentes são esperados após a exposição à alimentação, e os genes diferencialmente expressos podem ser usados como potenciais biomarcadores. Os resultados demonstraram que P. hawaiensis é um bom modelo para testes de toxicidade com um protocolo miniaturizado e o RNA-seq foi aplicado com sucesso para investigar as diferenças entre a exposição via alimentação com AgCl e Ag-NP e pode revelar potenciais biomarcadores. Além disso, este estudo desenvolveu protocolos e fornece informações moleculares que podem ser usadas em estudos futuros com P. hawaiensis. / Marine and estuarine ecosystems are the final destinations of many contaminants and ecotoxicology still faces the lack of model organisms for these environments. To develop and present a suitable test organism to be used in laboratory and field studies, methods for husbandry, ecotoxicity endpoints and biomarkers should be evaluated. The marine amphipod Parhyale hawaiensis is already a model for development and evolutionary studies, it is worldwide spread and easy to handle in the laboratory and so on an interesting model for ecotoxicology. The aim of this work is to standardize husbandry conditions, toxicity testing, and to develop molecular biomarkers in P. hawaiensis and apply them to evaluate the effect of metal nanoparticles in dietary exposure. The conditions for culturing were established in reconstituted seawater (30 ± 2 salinity), 24 ± 2 oC temperature, photoperiod 12-12 h light/dark, crushed coral as substrate, daily feeding with fish food, partial water exchange and constant aeration. Testing conditions comprised 96-well microplate, <7 days age organisms, one organism in each well containing 200 &#181;L of exposure media for 96 h. Zinc, copper, silver, cadmium and ammonia were selected as toxicants and sensitivity of P. hawaiensis were within the range of other marine amphipods. Gene expression conditions for P. hawaiensis were develop for the reference gene 18S, two metallothioneins (MT) and two glutathione-S-transferase (GST). However, selected target genes, analyzed by RT-qPCR, were not successful to detect differences in our treatment conditions. Thus, global gene expression analysis, RNA-seq, was conducted with adult organisms fed for 7 days to a contaminated food with silver nanoparticles (Ag-NPs), AgCl and control. Results highlighted differences between males and female\'s toxicity responses as well as AgCl and Ag-NP treatments. Comparison analysis revealed common genes expressed in both Ag forms, but also showed differences in Ag-NP treatment with both up and down regulated genes. Analysis are underway to better understand toxicity responses pathways. These results anticipate different responses to both Ag forms investigated, therefore different adverse effects are expected after feeding exposure to AgCl and Ag-NP and significative genes can be used as potential biomarkers. Our results demonstrated that P. hawaiensis is a good model for ecotoxicity tests with a miniaturized protocol and RNA-seq were successful applied to investigate the differences between AgCl and Ag-NP feeding exposure and could reveal promising biomarkers. Moreover, this study developed protocols and provide molecular information that can be used in future studies with P. hawaiensis.

Nanopartículas de prata (AgNPs) vêm ganhando grande importância na prevenção contra infecções, principalmente pela sua incorporação em dispositivos médicos, tecidos entre outros produtos relacionados. Estudos prévios indicaram que partículas de menores tamanhos têm ação biocida mais efetiva. Nesse contexto, a separação granulométrica pode ser explorada para se obter partículas com as dimensões desejadas. A centrifugação foi escolhida, entre outros métodos de separação granulométrica, por sua possibilidade de ressuspensão das AgNPs após a separação, pela facilidade do aumento de escala e baixa complexidade. Um modelo do perfil de concentração (em uma cela centrífuga) de AgNPs em função do tempo e intensidade de centrifugação foi proposto a partir da Equação de Lamm, considerando a difusividade Browniana e o coeficiente de Svedberg calculado a partir do balanço de forças. A equação foi resolvida com a solução semi-analítica proposta inicialmente por Fujita (1962) e melhorada pelos alemães Joachim Behlke e Otto Ristau (2001). O desempenho da centrifugação de AgNPs sintetizadas pelo Método Turkevich foi avaliado analisando-se as partículas sobrenadantes por DLS, UV-Vis e microscopia eletrônica e comparando com o modelo proposto. O modelo previu razoavelmente a concentração total de partículas e a sua distribuição de tamanhos em diferentes tempos de centrifugação para forças de campo baixas (força centrífuga relativa até 1480). Para maiores forças de campo, houve uma sedimentação mais acelerada que a prevista pelo modelo. / Silver nanoparticles (AgNPs) have gained great importance in preventing infections, especially by their incorporation into medical devices, tissues and related products. Previous studies indicated that particles of smaller sizes have more effective biocidal action. In this context, granulometric separation may be explored to deliver nanoparticles of desired size. Centrifugation was chosen, among other particle size separation methods, for their ability to resuspend the AgNPs after separation, for the ease of scale-up and low complexity. A profile concentration model (in the centrifuge cell) of AgNPs as a function of time and intensity of centrifugation has been proposed from the Lamm equation, considering the Svedberg and Brownian diffusivity coefficients calculated from a force balance. The equation was solved with the semi-analytical solution proposed initially by Fujita (1962) and improved by the Germans Joachim Behlke and Otto Ristau (2001). The centrifugation performance of AgNPs synthesized by the Turkevich method was evaluated by analyzing the supernatant particles by DLS, UV-Vis and electron microscopy and comparing to the proposed model. The model reasonably predicted the total concentration of particles and their size distribution at different centrifugation times at low field strengths (1480 RCF). For higher field strengths, the sedimentation was faster than expected by the model.

Orientador: Maria do Carmo Gonçalves / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-25T19:34:46Z (GMT). No. of bitstreams: 1 DeFarias_MarceloAlexandre_D.pdf: 6813625 bytes, checksum: 734f14c74ec0690013d0549463dd8b09 (MD5) Previous issue date: 2014 / Resumo: O principal objetivo desta tese consiste na síntese de nanopartículas de prata e de sílica e sua posterior modificação de superfície. Visando atribuir um caráter hidrofóbico às nanopartículas, sintetizou-se o copolímero em bloco de poliestireno-bloco-poli(N-vinil-2-pirrolidona) (PS-b-PVP), via técnicas de polimerização radicalar controlada. A metodologia utilizada produziu copolímeros de PS-b-PVP com baixa polidispersidade e massa molar controlada, capazes de estabilizar a emulsão de água em clorofórmio. Nanopartículas de prata (AgNP) produzidas à temperatura ambiente exibiram diâmetros médios de 8 e 11 nm, utilizando PVP ou PS-b-PVP como agentes estabilizantes, respectivamente. Nanopartículas de sílica (SiO2NP) foram sintetizadas pelo método de Stöber e apresentaram diâmetros aproximados de 40, 80 e 150 nm. Também foram utilizadas SiO2NP comerciais, que apresentaram duas populações com diâmetros médios de 12 e 25 nm, além de aglomerados. Por este motivo, a adsorção de PVP ou de PS-b-PVP à superfície das SiO2NP só foi caracterizada adequadamente com as SiO2NP sintetizadas. Mapas elementares de carbono, obtidos por imagem espectroscópica de elétrons (ESI-TEM) de dispersões de AgNP ou SiO2NP, mostraram que ambas as nanopartículas tiveram sua superfície modificada por PVP ou PS-b-PVP. Adicionalmente, as nanopartículas foram incorporadas em matrizes de poliestireno-bloco-polibutadieno-bloco-poliestireno (SBS) para produção de nanocompósitos. Observou-se por espalhamento de raios X de baixo ângulo (SAXS) e microscopia eletrônica de transmissão (TEM) que filmes de SBS puros apresentaram elevado grau de auto-organização da microestrutura de cilindros empacotados hexagonalmente, devido às condições de tratamento térmico e de casting utilizadas. Os nanocompósitos contendo SiO2NP apresentaram o mesmo comportamento termomecânico que a matriz de SBS, embora perturbações na microestrutura da matriz tenham sido observadas. Imagens de microscopia de força Kelvin mostraram a presença de domínios com potenciais elétricos distintos, indicando a ocorrência de uma forte interação entre o SBS e as AgNP/PS-b-PVP. Análises de DMA e TEM sugeriram que as AgNP/PS-b-PVP apresentaram maior interação com os cilindros de PS da matriz de SBS, em comparação com as AgNP/PVP. O conjunto de resultados obtidos neste trabalho indica que foi possível modificar superficialmente as nanopartículas inorgânicas empregando o copolímero PS-b-PVP / Abstract: The main objective of this thesis is the synthesis and surface modification of silver and silica nanoparticles. Aiming towards a hydrophobic character for the nanoparticles, the polystyrene-block-poly(N-vinyl-2-pyrrolidone) (PS-b-PVP) amphiphilic block copolymer was synthesized by controlled radical techniques. This methodology produced PS-b-PVP block copolymers with low polydispersity and controlled molar mass, which are able to stabilize water/chloroform emulsions. Silver nanoparticles (AgNP) produced at room temperature showed 8 and 11 nm mean diameters when using PVP or PS-b-PVP, respectively. Silica nanoparticles (SiO2NP) synthesized by Stöber method showed 40, 80 and 150 nm mean diameters. Commercial SiO2NP was also used and presented two populations with mean diameters of 12 and 25 nm, as well as agglomerates. The adequate characterization of PVP or PS-b-PVP adsorption onto SiO2NP surfaces was only possible using the SiO2NP obtained by the Stöber method. Carbon maps, obtained by electron spectroscopic imaging (ESI-TEM) showed that AgNP and SiO2NP were both surface modified by PVP or PS-b-PVP. Additionally, the nanoparticles were incorporated into polystyrene-block-polybutadiene-block-polystyrene (SBS) matrices to produce nanocomposites. Small angle X ray scattering (SAXS) and transmission electron microscopy (TEM) showed that SBS films presented a high self-assembly degree of the hexagonally packed cylinder due to annealing and casting conditions. Similar thermal-mechanical behavior was observed for the nanocomposites containing SiO2NP in relation to pure SBS, although perturbations in the SBS microstructure had also been observed. Kelvin force microscopy (KFM) showed domains with different electrical potentials, indicating a strong interaction between SBS and AgNP/PS-b-PVP. DMA and TEM analysis suggested that AgNP/PS-b-PVP interacted preferentially with SBS matrix PS cylinders. The results obtained in this work indicate that the inorganic nanoparticle surfaces were successfully modified by the PS-b-PVP copolymer / Doutorado / Físico-Química / Doutor em Ciências

Les travaux précédents du laboratoire Hubert Curien ont permis d’élaborer des films photosensibles Ag : TiO2 sur verre, support de marquage de motifs actualisables ou permanents. Une adaptation de ces travaux aux supports plastiques et papiers est ici proposée afin d’élargir les domaines d’application potentiels au marquage sécurité des produits par exemple. Des techniques d’élaboration de films Ag : TiO2 compatibles avec les substrats considérés ont donc été développées. Deux voies ont été envisagées. La première utilise la chimie du Sol-Gel combinée à la méthode EISA et des procédés de dépôt tel le spin-coating, le jet d’encre ou la flexographie pour former un film mésoporeux de TiO2 sur les supports. Des traitements basés sur une extraction par solvant ou un recuit infrarouge ont été imaginés afin de libérer la porosité du film sans dégradation du support. Pour réaliser les films sur papier, un sel d’argent est ajouté au Sol avant son dépôt. Dans les cas des films élaborés sur plastique, l’argent est incorporé par imprégnation du matériau dans une solution de sel d’argent. La deuxième voie d’élaboration propose de formuler une encre aqueuse jet d’encre de nanoparticules de TiO2 et d’ions argent, en adaptant la composition d’une suspension commerciale de TiO2 aux exigences du jet d’encre. Après dépôt, l’encre est séchée par recuit infrarouge. Le comportement photochromique sous expositions lumineuses UV et visible des différents films permet leur coloration et décoloration de façon réversible. Les films Sol-Gel déposés sur plastique peuvent également être support de photo-inscriptions permanentes générées par irradiation par une lumière visible d’une certaine intensité / Previous research conducted at laboratory Hubert Curien led to the development of photosensitive Ag: TiO2 films on glass as support for updatable or permanent patterns. An adaptation of this work to plastic and paper substrates is proposed here to broaden the possible application areas such as goods secure labeling, for example. Fabrication techniques of Ag: TiO2 films compatible with flexible substrates have been developed, using two different paths. The first uses the combination of Sol-Gel chemistry with the EISA method. Deposition processes such as spin coating, inkjet or flexographic printing are used to form a mesoporous film of TiO2 on substrates. Treatments based on solvent extraction or infrared annealing have been devised to release the porosity of the film without damaging the supports. Silver salt is either introduced into the titania pores by soaking the films into a silver salt solution or added to the sol before its coating. The second developed option proposes formulating an aqueous ink jet ink made of TiO2 nanoparticles and silver ions by adapting the composition of a commercial suspension of TiO2 with the requirements of the ink jet process. After printing, the ink is dried by infrared annealing. The photochromic behavior under UV and visible light exposures of fabricated films allows to get coloring and bleaching reversibly. Sol-Gel films coated on plastic can also be a support for permanent colored patterns realized by irradiation with a visible light of certain intensity

Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The development of polymers with the ability to transport and release drugs and bioactives has been growing rapidly because of their unique properties in protecting and enhancing solubilization of drugs in physiological media. In this work, polyesters derived from β-cyclodextrin were developed in two different systems: The former consisted of cyclodextrin cross-linked with citric acid and subsequently functionalized with glutamic acid. The latter was obtained from β-cyclodextrin esterified with gluthamic acid. The polyesters were characterized by Fourier Transform Infrared Spectroscopy (FTIR), Solid-state Cross-Polarization Magic Angle Spinning Carbon-13 Nuclear Magnetic Resonance (CP/MAS 13C–NMR) and Thermogravimetric Analysis (TGA) for the confirmation of the formation of polymers and the esterification of glutamic acid with β-cyclodextrin molecules. The formation of inclusion complex of the polymers in solution with orange methyl was evaluated to verify the activation of β-cyclodextrin cavities. The polymers were used as reducer and stabilizer in the synthesis of silver nanoparticles (AgNPs). The characterization of the AgNPs was performed by UV-Vis spectroscopy, and bands between 350-500 nm evidenced of the obtainment of silver nanoparticles. Analyses by transmission electron microscopy revealed AgNPs with spherical morphology and diameter around ~ 5 to ~60 nm, corroborating with the results of UV-Vis. / O desenvolvimento de polímero com a capacidade de transportar e liberar fármacos e compostos bioativos vem crescendo rapidamente, devido suas propriedades únicas em proteger e aumentar a solubilização em meios fisiológicos. Neste trabalho, foram desenvolvidos poliésteres derivado de β-ciclodextrina em dois sistemas distintos. O primeiro sistema foi obtido a partir de β-ciclodextrina reticulado com ácido cítrico e posteriormente funcionalizado com ácido glutâmico. O segundo foi a partir de β-ciclodextrina esterificada com ácido glutâmico. Os poliésteres foram caracterizados por espectroscopia na região do infravermelho com transformada de Fourier (FTIR), ressonância magnética nuclear no estado sólido sob polarização cruzada e rotação com ângulo mágico (RMN 13C CP/MAS) e análise termogravimétrica (TG), para a confirmação da formação dos polímeros e a esterificação do ácido glutâmico com as moléculas de β-ciclodextrina. A formação de complexo de inclusão dos polímeros com o alaranjado de metila foi avaliada em solução, comprovando-se a ativação das cavidades de β-ciclodextrina. Os polímeros foram testados quanto a habilidade de promover a formação/estabilização de nanopartículas de prata (AgNPs) a partir de nitrato de prata em solução. A caracterização das AgNPs foi realizada por espectroscopia de UV-Vis, e as bandas observadas entre 350-500 nm são evidências da presença de nanopartículas de prata. As imagens de microscopia eletrônica de transmissão revelaram AgNPs de morfologia esférica com diâmetro em torno de ~5 a ~60 nm, corroborando com os resultados de UV-Vis. / São Cristóvão, SE

Nanotrubičky oxidu titaničitého v průběhu posledních let nabyly na významu v poli biomedicíny. Jakožto biokompatibilní nanostrukturovaný povrch nachází potenciál pro své uplatnění především v oblasti implantačních aplikací. Teoretická část této práce je tak věnována různým přístupům pro syntézu TiO2 nanotrubiček, jejich modifikacím a aplikacím v biomedicíně. Experimentální část pak pojednává o nanotrubičkách oxidu titaničitého, které jsou připraveny z titanu metodou jednokrokové anodické oxidace v organickém elektrolytu. Jako výchozí materiály jsou používány křemíkové disky s naprášenou vrstvou titanu a titanové folie. Zprvu amorfní nanotrubičky jsou žíháním převedeny na svou krystalickou podobu, a následně modifikovány selenovými a stříbrnými nanočásticemi. Připravené struktury jsou zkoumány z hlediska povrchových vlastností a biologických interakcí s vybranými tkáňovými kulturami (MG-63, NIH-3T3) a bakteriemi (E. coli, P. aeruginosa, S. aureus). V závěru experimentální práce jsou stručně porovnány výsledky selenových a stříbrných nanočástic. Hlavním cílem této práce je rozšířit znalosti týkající se bio-rozhraní tvořeným adherentními buněčnými liniemi, bakteriálními buňkami a nanostrukturovaným povrchem tvořeným TiO2 nanotrubičkami dekorovanými selenovými a stříbrnými nanočásticemi.

Today, development of accurate early diagnosis of cancers thus became the number one challenge of medicine during the 21st century as the current techniques relies on imaging methods that suffer from low sensitivity and misdiagnosis.For these reasons, in this work, we aimed at developing a no-wash colorimetric biosensor for the detection of the oncoprotein Mdm2. Indeed, abnormal levels of Mdm2 could be related to the early formation of tumors.This thesis is devoted to the conception of a no-wash colorimetric biosensor for the detection of the oncoprotein Mdm2. This work can be divided in four parts:(i) The detection strategy and the design of the recognition elements (Chapter I).(ii) The conjugation of gold nanoparticles with the recognition elements (Chapter II, III, IV, V and VI).(iii) The modification of the metallic core of the nanoparticles (Chapter VII).(iv) The use of the optimized biosensor for the detection of Mdm2 (Chapter VIII).In the first part, we investigated the sensing strategy. An aggregation-based assay with plasmonic nanoparticles was selected, as the detection signal is a change of color of the suspension that can be observed to the naked eye or by UV-Vis spectroscopy. We designed the recognition elements, two peptide aptamers coming from endogenous proteins p53 and p14, and we grafted them separately on two batches of gold nanoparticles (AuNPs) via thiol end-groups. We used these latter for the detection of various concentrations of Mdm2 in buffer using our dual-trapping strategy with these two batches of functionalized AuNPs. We demonstrated that both peptides are able to interact with Mdm2 even after grafting onto the particles and that this detection strategy is highly specific. However, this first sensor presented some drawbacks, such as a poor colloidal stability of the AuNPs and a limited dynamic range.With the aim to encompass these issues we investigated, in the second part of this thesis, alternative strategies to conjugate the peptides to the particles. We investigated the functionalization of the particles with stabilizing ligands such as thiolated poly(ethyleneglycol) (HS-PEG). We first studied their simultaneous grafting with the peptides on the AuNPs. We observed that grafting HS-PEGs and peptides side-by-side allowed to control the density of peptides conjugated to the AuNPS and increased drastically the stability of the particles. However, the detection of Mdm2 was strongly hindered by the presence of PEG on the particles carrying the p14 peptide. In a second step, we investigated the conjugation of peptides on the top of a PEG layer carrying functional groups (HS-PEG-X where X is a carboxylate or an alkyne). AuNPs were first functionalized with mixtures of HS-PEG and HS-PEG-X, and the peptides were conjugated to the functional groups via amide bond formation or CuAAC coupling in a second step. However, we noticed that it was not possible to control the composition of the mixed layer of PEGs and thus the peptide grafting density.Due to the lack of recognized protocols in the literature for (i) the determination of the chemical and colloidal stabilities of AuNPs and (ii) the determination of the proportion of different ligands in the organic coating of the particles, we developed two interesting tools. The first one was a convenient method allowing to evaluate by UV-Vis spectroscopy the efficiency of the citrate exchange process using thiol-, alkyne- or diazonium-ligands from gold nanoparticles synthesized via a Turkevich method. The second protocol was a method allowing to quantify the proportion of two HS-PEGs ligands grafted in mixtures onto gold nanoparticles via 1H NMR spectroscopy.As we couldn’t find conditions in which the proportion of multiple thiolated ligands can be controlled on AuNPs, we decided to investigate another functionalization strategy based on the use of calix4arene-diazonium salts.We first studied the grafting on AuNPs of calixarenes bearing four PEG chains at the level of their small rim, one ended by a carboxylic acid and three by a methoxy group. The calixarene layer allowed to obtain AuNPs covered by a very dense PEG shell (with more PEG chains/nm2 that what was obtained previously with thiol anchoring). In addition to that, this PEG shell was strongly anchored to the AuNPs, conferring them a very high colloidal and chemical robustness. We then combined the grafting of this calixarene with the grafting of another non-functional calixarene, bearing four PEG chains ended by a methoxy group, and we quantified the conjugation capacity of such particles by amide bond formation. We demonstrated that this strategy allows to (i) increase drastically the stability of the AuNPs and (ii) control the proportion of peptide conjugated at their surface. Finally, we showed that calixarene-coated AuNPs to which to the p53 and p14 peptides have been conjugated could be used to detect Mdm2.With the evidence that the peptide conjugation density could be controlled using calixarene-coated AuNPs, we investigated the simultaneous grafting of two functional calixarenes on particles: one bearing four carboxylic acids groups and one bearing four PEG chains ended by alkyne groups. We optimized the grafting of these calixarenes in mixed layers on the AuNPs as well as their conjugation. We demonstrated that the grafting of two functional calixarenes led to the production of bi-functional AuNPs, capable of conjugation with two molecules via two distinct chemistries.In the third part, we optimized the composition of the metallic core of the biosensor. As it is well known that silver nanoparticles express better optical properties than gold nanoparticles of the same size, we aimed to incorporate silver nanoparticles (AgNPs) in the biosensor. This was a true challenge due to the intrinsic low chemical stability of silver nanoparticles that greatly limits their use in IVD. For this purpose, we developed an innovative in situ synthesis of silver nanoparticles in the presence of the calixarene-diazonium salts. After optimization of the synthesis, we observed that calixarenes bearing four carboxylic acids groups at the level of their small rim allowed the production of ultra-stable silver nanoparticles to which biomolecules can easily be conjugated. This in situ synthesis procedure even allowed us to produce alloy nanoparticles, with metallic cores whose composition could easily be tuned from pure silver to silver/gold alloys or pure gold. With this synthesis, the composition of the organic layer could also be easily tuned by using mixtures of calixarenes-diazonium salts.Finally, in the last part, we investigated the detection of Mdm2 with the optimized version of the biosensor, i.e. silver nanoparticles coated by a calixarene layer to which the p53 and p14 peptides were conjugated. With this novel class of nanoparticles, we could encompass the two initial drawbacks of the initial sensor. First, we were able to detect Mdm2 with a wider detection range and a lower limit. Secondly, the particles were sufficiently stable and robust to be dispersed in physiological fluids and we could detect Mdm2 in human serum without interference. / Doctorat en Sciences de l'ingénieur et technologie / info:eu-repo/semantics/nonPublished

Tato diplomová práce se zabývá studiem plasmonicky navýšené absorpce vedoucí ke zlepšení účinnosti organických solárních článků. K navýšení absorpce světla ve fotoaktivní vrstvě jsou použity koloidní nanočástice stříbra. Rozptyl světla z nanočástic střibra do fotoaktivní vrstvy představuje jedno z možných řešení, jak navýšit celkovou účinnost fotovoltaických zařízení. Simulace elektromagnetických jevů jsou pro statické podmínky prováděny v softwaru Lumerical (Lumerical Solutions, Inc.). Je zkoumána absorpce ve fotoaktivní vrstvě v závislosti na konfiguraci stříbrných nanočástic. Simulace potvrzují, že ve fotoaktivní vrstvě, která je modifikovaná nanočásticemi stříbra, dochází k navýšení absorpce. Abychom potvrdili výsledky simulací, bylo vyrobeno několik stříbrem modifikovaných vzorků. Vzorky byly pro tento druh experimentu připravené z poly(3-hexyltiofenu):[6,6]-fenyl-C61-butyric-acid-metyl esteru. U některých, nanočásticemi stříbra modifikovaných, vzorků dochází k navýšení tvorby excitonů, v důsledku čehož je pozorován nárůst fotoproudu. V této práci je zahrnut teoretický i experimentální přístup k dané problematice.

Les premières photographies couleurs produites par Edmond Becquerel au Muséum d’Histoire naturelle à Paris en 1848 ont suscité un débat scientifique intense tout au long du XIXème siècle. La question de l’origine des couleurs de ces photographies a vu s’affronter les partisans d’une hypothèse pigmentaire et ceux d’une hypothèse interférentielle. Au cours du XXème siècle, cette question n’a pas été définitivement tranchée. Ce travail doctoral se propose d’appréhender la nature de ces images dites « photochromatiques » par une approche expérimentale. Nous nous sommes dans un premier temps intéressés aux étapes de sensibilisation du plaqué argent et d’exposition de la surface sensible aux rayonnements visibles, dans le but de répliquer le procédé Becquerel et de comprendre la photosensibilité de ces images. Les propriétés optiques des couches sensibles et colorées ont été caractérisées en spectroscopie UV-visible et mises en relation avec leurs compositions chimiques (étudiées en XAS) et leurs morphologies, de l’échelle micro- à nanométrique (étudiées en MEB et TEM). La grande sensibilité des échantillons aux faisceaux photoniques (UV-visible et rayons X) et électroniques a imposé un développement méthodologique afin de comprendre et de limiter les effets de sonde. Les résultats de cette recherche permettent de réfuter l’hypothèse de phénomènes interférentiels comme origine des couleurs des images photochromatiques, hypothèse qui prévaut dans la littérature depuis la fin du XIXème siècle. Les couches sensibles et colorées sont constituées de grains de chlorure d’argent micrométriques décorés de nanoparticules d’argent. Ces nanoparticules d’argent, que nous avons étudiées en EELS low loss, sont responsables de l’absorption dans le visible des couches sensibles et colorées par résonance de plasmons de surface. Ceci nous conduit à proposer une origine plasmonique aux couleurs des images photochromatiques, hypothèse discutée dans cette thèse. / The first colour photographs were produced in 1848 by Edmond Becquerel at the Museum of natural History in Paris. The origin of their colours motivated an intense debate between the scientists during the XIXth century. Two main hypotheses were proposed, namely a pigmentary hypothesis and an interferential hypothesis. Nowadays the question of the colours of those photochromatic images is still not settled and this doctoral research aims at addressing it by an experimental approach. We first studied the sensitizing of the silver plate and the exposure of the sensitive surface to light in order to replicate the Becquerel process and to gain information on the photosensitivity of these images. The optical properties of the sensitized and coloured layers were characterized by UV-visible spectroscopy and related to their chemical composition (studied by XAS) and their morphologies, from the micro to the nanoscale (studied by SEM and TEM). A methodological development was necessary to understand and overcome the sensitivity of the samples to photonic beams (UV-visible and X-rays) and electronic beams. The results allow us to reject the interferential hypothesis, which prevailed since the end of the XIXth century. The sensitized and coloured layers consist in micrometric silver chloride grains decorated by silver nanoparticles. Besides, these nanoparticles, which we investigated by low loss EELS, are responsible for the visible absorption of sensitized and coloured layers through surface plasmon resonance. We suggest that the photochromatic images colours have a plasmonic origin and hereby discuss this hypothesis.

Les nanoparticules métalliques (NPs) présentent des propriétés optiques (PO) uniques provenant de l’oscillation collective de leurs électrons. Cet effet se traduit par l'émergence d'une bande plasmon dont les caractéristiques peuvent être modulées par la taille, la forme, la nature des NPs et le milieu hôte. Il existe de nombreuses méthodes pour la préparation de ces NPs, l'une d'entre elles est l'ablation laser en milieu liquide (ALML). Cette technique offre certains avantages comme la simplicité, l’adaptabilité et des NPs dépourvues de contamination. Ses principaux inconvénients sont la productivité et le contrôle de la taille et de la forme des NPs. Ce travail est consacré à l'élaboration de NPs d’Ag par l’ALML et à l'étude théorique de leurs PO. Nous donnons dans ce manuscrit, les résultats de l'optimisation des paramètres d'élaboration conduisant à l'obtention de distributions en NPs reproductibles et contrôlées. Les PO de ces NPs sont mesurées et comparées à des modèles physiques spécifiques basés sur la théorie des milieux effectifs (EMT). L'EMT, telle que le modèle de Maxwell-Garnett, permet de décrire les PO de NPs monodisperses. Cependant, les voies de préparation classiques conduisent inévitablement vers des NPs montrant une distribution de forme et de taille qui induit des changements drastiques sur leurs PO. Le modèle SDEMT est proposée pour le calcul de la fonction diélectrique effective et du coefficient d'absorption de solutions colloïdales de NPs métalliques. Contrairement à Maxwell-Garnett, ce modèle donne une meilleure description des spectres d'absorption et d’ellipsométrie mesurés sur des échantillons contenant des NPs d’Ag et d’Au / Metal nanoparticles (NPs) exhibit unique optical properties (OP) coming from the collective oscillations of their electrons. This effect is translated by the emergence of a band of plasmon, the characteristics which can be modulated by the size, the shape and the nature of the NPs as well as by the environment of the host. There are many methods for the preparation of NPs, and one of them is the pulsed-laser ablation in liquid (PLAL). This technique offers some advantages such as simplicity, versatility and surface NP without contamination (reducing agent residues and/or stabilizers). Its main drawbacks are the lacks of productivity and control of the NP size and shape. This work is devoted to elaboration of Ag NPs by PLAL and theoretical investigation of their OP. We give here the results about the optimization of elaboration parameters leading to obtaining reproducible and controlled distributions of Ag NPs. The OP of these NPs are measured and compared to specific physical models based on the effective medium theory (EMT). Classical EMT such as Maxwell Garnett approximation allows describing the OP of monodisperse NPs. However, conventional preparation routes unavoidably conduct to NPs showing a shape and a size distribution which induces drastic changes in the OP. A SDEMT model which considers the shape dispersion is proposed for the calculation of the effective dielectric function and absorption coefficient of colloidal solution of metal NPs in water. Contrary to the conventional theory, this model gives a better description of the measured absorption and ellispometry spectra of sample containing Ag and Au NPs