A coleta eficiente da radiação através de processos de transferência de energia de ressonância Förster (FRET, por sua sigla em inglês) constitui uma forma atraente de melhorar aplicações nanotecnológicas que envolvam o uso da radiação solar como fonte de energia. Sistemas constituídos de zeólita L funcionalizada em sua superfície com ftalocianina (PC) foram estudados visando a inativação de patógenos mediante o uso de luz solar natural. No entanto, na velocidade de fotoinativação observou-se uma dependência com a intensidade da radiação que foi relacionada com a absorção ineficiente de PC na região transversal de 400-650 nm da irradiância solar. Visando uma aplicação nesses sistemas, neste trabalho estudou-se a transferência de energia tipo Förster utilizando os corantes catiônicos Pironina Y (PyY) e Tionina (Th) encapsulados na zeólita L que se caracterizam por absorver na região espectral visível onde a absorção de PC é limitada. Os corantes foram inseridos no interior dos canais da zeólita L por um processo de troca iônica e a distância de separação entre estes foi variada com o carregamento das amostras (% de moléculas inseridas em relação ao espaço disponível na zeólita). Foram preparadas amostras com a inserção de PyY e Th nos canais da zeólita L, individualmente, bem como, amostras dos dois corantes misturados aleatoriamente na razão doador:receptor 1,0:1,4. A transferência de energia foi estudada mediante a detecção da emissão sensibilizada do receptor Th quando o doador PyY foi seletivamente excitado nas amostras de PyY-Th-zeólita L e, através da comparação dos tempos de vida de fluorescência do doador na presença e na ausência do receptor. O conjunto de resultados a diferentes carregamentos sugere a FRET nas amostras testadas, no entanto, devido à baixa emissão de Th detectada, o material preparado não seria adequado para a transferência de energia ao PC. / The efficient harvesting of solar energy through Förster Resonance Energy Transfer processes (FRET) is an attractive way to improve nanotechnology applications involving the use of solar radiation as energy source. Systems based on zeolite L functionalized on its surface with phthalocyanine (PC) have been studied to inactivate pathogens by natural sunlight. However, there was observed a dependency between the photoinactivation rate and the radiation intensity which was related to the inefficient absorption of PC in the vertical cross-section of 400-650 nm of the solar spectral irradiance. Aimed at applying in these systems, we studied the Förster type energy transfer between the cationic dyes Pyronin Y (PyY) and Thionin (Th) encapsulated into the zeolite L which are characterized by absorbing in the visible spectral region where the absorption of PC is limited. The dyes were inserted inside the zeolite L channels through ion exchange process, and the separation distance between them was tuned with the sample loading (% of inserted molecules in relation to the available space in the zeolite). Different samples were prepared with one and two randomly mixed dyes keeping the donor:acceptor ratio in 1.0:1.4. The energy transfer was studied by detection of the acceptor sensitized emission when the donor PyY was selectively excited in PyY-Th-zeolite L samples, and by comparison of the donor fluorescence lifetimes in the presence and absence of the acceptor. The set of results in different loadings suggests FRET in the samples tested, however, due to the low detected emission of Th the prepared material would not be appropriate for energy transfer to PC.

A coleta eficiente da radiação através de processos de transferência de energia de ressonância Förster (FRET, por sua sigla em inglês) constitui uma forma atraente de melhorar aplicações nanotecnológicas que envolvam o uso da radiação solar como fonte de energia. Sistemas constituídos de zeólita L funcionalizada em sua superfície com ftalocianina (PC) foram estudados visando a inativação de patógenos mediante o uso de luz solar natural. No entanto, na velocidade de fotoinativação observou-se uma dependência com a intensidade da radiação que foi relacionada com a absorção ineficiente de PC na região transversal de 400-650 nm da irradiância solar. Visando uma aplicação nesses sistemas, neste trabalho estudou-se a transferência de energia tipo Förster utilizando os corantes catiônicos Pironina Y (PyY) e Tionina (Th) encapsulados na zeólita L que se caracterizam por absorver na região espectral visível onde a absorção de PC é limitada. Os corantes foram inseridos no interior dos canais da zeólita L por um processo de troca iônica e a distância de separação entre estes foi variada com o carregamento das amostras (% de moléculas inseridas em relação ao espaço disponível na zeólita). Foram preparadas amostras com a inserção de PyY e Th nos canais da zeólita L, individualmente, bem como, amostras dos dois corantes misturados aleatoriamente na razão doador:receptor 1,0:1,4. A transferência de energia foi estudada mediante a detecção da emissão sensibilizada do receptor Th quando o doador PyY foi seletivamente excitado nas amostras de PyY-Th-zeólita L e, através da comparação dos tempos de vida de fluorescência do doador na presença e na ausência do receptor. O conjunto de resultados a diferentes carregamentos sugere a FRET nas amostras testadas, no entanto, devido à baixa emissão de Th detectada, o material preparado não seria adequado para a transferência de energia ao PC. / The efficient harvesting of solar energy through Förster Resonance Energy Transfer processes (FRET) is an attractive way to improve nanotechnology applications involving the use of solar radiation as energy source. Systems based on zeolite L functionalized on its surface with phthalocyanine (PC) have been studied to inactivate pathogens by natural sunlight. However, there was observed a dependency between the photoinactivation rate and the radiation intensity which was related to the inefficient absorption of PC in the vertical cross-section of 400-650 nm of the solar spectral irradiance. Aimed at applying in these systems, we studied the Förster type energy transfer between the cationic dyes Pyronin Y (PyY) and Thionin (Th) encapsulated into the zeolite L which are characterized by absorbing in the visible spectral region where the absorption of PC is limited. The dyes were inserted inside the zeolite L channels through ion exchange process, and the separation distance between them was tuned with the sample loading (% of inserted molecules in relation to the available space in the zeolite). Different samples were prepared with one and two randomly mixed dyes keeping the donor:acceptor ratio in 1.0:1.4. The energy transfer was studied by detection of the acceptor sensitized emission when the donor PyY was selectively excited in PyY-Th-zeolite L samples, and by comparison of the donor fluorescence lifetimes in the presence and absence of the acceptor. The set of results in different loadings suggests FRET in the samples tested, however, due to the low detected emission of Th the prepared material would not be appropriate for energy transfer to PC.

L'imagerie de fluorescence s'est particulièrement développée au fil de ces dernières décennies notamment pour l'exploration des systèmes biologiques. De nombreux développements portant à la fois sur les instruments d'analyse et les agents de contraste (sondes) ont été entrepris pour améliorer cette technique de bioanalyse. Mes travaux de thèse avaient pour but d'explorer diverses plateformes moléculaires pro-fluorescentes pouvant générer, sous l'effet d'un stimulus biologique / chimique, un fluorophore organique. Cette approche de synthèse chimique in-situ met en jeu des réactions domino caractérisées par la formation et la rupture de liaisons covalentes. Ce processus devrait ouvrir la voie à des sondes fluorogéniques possédant des rapports signal sur bruit élevés. Cette étude avait aussi pour but d'étudier la synthèse in-situ de fluorophores déclenchée par plusieurs stimuli afin de concevoir des portes logiques moléculaires de type "AND", dans un but de détection "multi-analytes". Pour établir la preuve de principe, la formation d'un coeur 7-hydroxy-2-iminocoumarine fluorescent déclenchée par divers couples d'enzymes (hydrolase / nitroréductase) a été étudiée. L'ensemble de ces travaux sont décrits dans le chapitre I de ce manuscrit. Par la suite et comme présenté dans les chapitres II et III, notre intérêt s'est porté sur le développement des plateformes fluorogéniques libérant des fluorophores ayant des maxima d'absorption / émission décalés vers le rouge en comparaison de ceux des coumarines. Une première réalisation a concerné la conception d'un précurseur "cagé" agissant comme sonde à peptidases ou nitroréductase par libération d'un dérivé pyronine. Des travaux portant sur la formation in-situ d'une benzophénoxazine sont également en cours et sont présentés au chapitre III. Un dernier aspect de ce travail a concerné la chimie d'une nouvelle famille de fluorophores proche-IR (les hybrides dihydroxanthène-hémicyanine) dont la formation in-situ pourra être envisagée pour des applications in-vivo. / Fluorescence imaging is a growing field of biology over the past decades. Intensive works mainly focused on instrumental developments and chemistry of contrast agents (probes), were already done to improve such bioanalytical technique. The main goal of this Ph. D. thesis was to explore various fluorogenic molecular platforms responsive to various (bio)chemical stimuli and capable of releasing organic fluorophores in the biological medium to analyze. This approach named "in-situsynthesis" is based on domino reactions belonging to the repertoire of "covalent chemistry", triggered by the target (bio)analyte. This kind of process should provide advanced fluorogenic probes with high signal-to-noise ratio. Another purpose of this work was to investigate some dualtriggering events to access to fluorogenic molecules acting as "AND-type" molecular logic gates for dual-analytes detection applications. To establish this approach, the formation of highly fluorescent7-hydroxy-2-iminocoumarin scaffolds triggered by several enzyme pairs (hydrolase and nitroreductase) was studied and now described in the first chapter of this manuscript. The second part of this work, described in chapters II and III, was devoted to the development of original "caged" precursors able to release fluorophores whose absorption / emission maxima are dramatically redshifted compared to those of coumarin derivatives. The first achievement concerned the detection of protease or nitroreductase activities through in-situ formation of a pyronin scaffold. Further works, currently in progress, are focused on "caged" precursors whose the dual reaction with a model protease should lead to the release of a benzophenoxazine derivative. Finally, some chemistry aspects related to an emerging and promising class of NIR fluorophores (dihydroxanthene-hemicyanine hybrids) are presented and the opportunity to form them in biological media, upon enzymatic triggering is also discussed.