Дисертації з теми "Imagerie multiplexé en fluorescence"
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Chouket, Raja. "New dimensions for multiplexed fluorescence imaging." Electronic Thesis or Diss., Sorbonne université, 2019. http://www.theses.fr/2019SORUS606.
Повний текст джерелаOur research group had previously developed the OPIOM protocols for fluorescence imaging. By exploiting their cross sections of fluorescence photoswiching, OPIOM can selectively extract the response of reversibly photoswitchable fluorophores (RSFs) in the presence of spectrally interfering fluorophores. However, OPIOM allowed us to discriminate only 3 spectrally similar reversibly photoswitchable fluorescent proteins (RSFPs). The goal of this PhD was to augment this number. To reach this goal, a new automated instrumental setup called photoswichometer was first developed to express and screen the rich photochemical signature of 22 RSFPs by analyzing their fluorescence response to light jumps with intensities covering 5 orders of magnitude. This signature has been first exploited in a new fluorescence imaging protocol called HIGHLIGHT, which capitalized on OPIOM and further improved its selectivity. In HIGHLIGHT, the RSFs are submitted to harmonic light modulation and their contribution to the overall fluorescence emission signals is selectively retrieved from exploiting their singular non-linear response under optimized conditions. HIGHLIGHT has been implemented to image RSFPs in cells without interference of autofluorescence, to perform multiplexed imaging of 3 RSFPs which could not be discriminated with OPIOM, and used for its intrinsic optical sectioning. The RSF signature has been then used in a second fluorescence imaging protocol called LIGHTNING. In contrast to OPIOM and HIGHLIGHT which exploit the cross sections of fluorescence photoswitching in a steady-state regime of low light intensity, LIGHTNING exploits the transient time fluorescence response of RSFs under multiple illuminations involving various ranges of light intensities for RSF discrimination. Thus, LIGHTNING allowed us to improve the multiplexing degree of dynamic contrast in fluorescence imaging up to 20 RSFP among 22 studied RSFPs
Deiss, Frédérique. "Développement de réseaux multiplexés de biocapteurs électrochimiques." Thesis, Bordeaux 1, 2009. http://www.theses.fr/2009BOR13883/document.
Повний текст джерелаThis work presents the development of optoelectrochemical micro- and nanosensor arrays for bioanalytical applications. These platforms respond to the growing need in research and diagnostic for tools allowing multiple and simultaneous analysis in small-volume samples. These new high density biochips are made from coherent optical fiber bundles: one face is micro- or nanostructured by chemical etching and then functionnalized with biological probes. The first biochip is a fluorescent DNA nanosensor array where probes have been immobilized by electrodeposition of a polypyrrole thin film. The detection of the hybridization is remotely performed through the imaging fiber. Different probes were succesfully addressed onto the same nanostructured array thanks to electrochemical cantilevers. The second biochip allows multiplexed sandwich immunoassays using electrochimiluminescent imaging resolved at the single bead level. In particular, the development of this new readout mechanism allows extending electrochemiluminescent detection for multiplexed immunoassays. Design and implementations of both platforms take advantages of different physical and chemical techniques, especially electrochemical, to obtain biochips with a great potential through high density and high multiplexing level
Valisa, Paolo. "Imagerie spectrale en raman et fluorescence : developpement et applications (doctorat : pharmacie)." Reims, 1997. http://www.theses.fr/1997REIMP204.
Повний текст джерелаProvencher, David. "Imagerie 3D de l'anatomie interne d'une souris par dynamique de fluorescence." Mémoire, Université de Sherbrooke, 2012. http://hdl.handle.net/11143/6205.
Повний текст джерелаMaunoury, Vincent. "Imagerie de fluorescence ph dependante en oncologie : approche experimentale chez l'animal." Lille 2, 1994. http://www.theses.fr/1994LIL2P264.
Повний текст джерелаLauffer, Mathieu. "Reconnaissance d'organismes aquatiques envahissants par traitement d'image et imagerie de fluorescence." Thesis, CentraleSupélec, 2015. http://www.theses.fr/2015SUPL0024.
Повний текст джерелаPhytoplankton plays a fundamental role in the living world. It is a dioxygen generator and the most important carbon dioxide fixer on Earth. However, under certain conditions, its development may become so excessive that it could be harmful to other vegetal and animal aquatic life in the water ponds in which it grows: it is the “hyper eutrophication” phenomenon. Such a situation leads to dramatic consequences on environment due to the difficulties that arise for photosynthesis and gas exchanges of other plant species. At the extreme limit, this can cause the death of the whole aquatic ecosystem. It appears therefore essential to strengthen the vigilance on controlling the proliferation of plankton and toxins with the necessity of risk evaluation. In a first approach, the recognition and the identification of aquatic organisms, necessary for such a control, are usually performed only by specialists algologists from microscopic observations. Nevertheless, in certain circumstances, it may be useful to dispose of an automatic recognition system to improve the monitoring of high-risk water ponds and optimize human intervention of specialists algologists. The development of such an automatic system of recognition of aquatics organism is more and more considered.In order to identify aquatic organisms, an original optical microscopy set up was developed in which the incident and emitted light beams are filtered in wavelengths. Such a set up enables the acquisition of classical microscopic images and microscopic images of fluorescence emission of the vegetal material under different illumination. These different images are then analyzed and processed by two algorithms of segmentation to collect characteristics data of the vegetals morphology and pigments compositions useful thereafter for their automatic recognition. Finally, all these different characteristic parameters linked to morphology and fluorescence emission of the vegetal species are collected to build a database useful for automatic optical recognition
Piraux, Hélène. "Nanoparticules magnétiques : Vectorisation par la voie principale d'acquisition du fer : Elaboration et physico-chimie." Paris 7, 2014. http://www.theses.fr/2014PA077219.
Повний текст джерелаDue to their magnetic properties, iron oxide nanoparticles are widely used for their significant role in the diagnostic and treatment of cancer. One of the main challenges of magnetic nanoparticles in nanomedicine is their effective targeting, which consists of their fast and precis( delivery directly into destination cells. Transferrin is one of the two proteins involved in the major iron acquisition pathway. Indeed, via its interaction with Receptor 1, transferrin crosses the plasma membrane within few minutes by receptor-mediated endocytosis. Furthermore, the overexpression of transferrin-receptor 1 in cancer tells enhances transferrin internalization making it a perfect Trojan horse for nanoparticles delivery. In this work, 3 different sizes of maghemite nanoparticles (5, 10 and 15 nm) were synthetized by the polyol method, coated with 3-aminopropyltriethoxysilane and coupled to transferrin. For each size, the ratio of transferrin per nanoparticle was determined and the interaction in vitro with the receptor was investigated. Then a comparative study of internalization was conducted in cellulo between raw and grafted to transferrin nanoparticles. The efficiency of the targeting was analyzed by magnetophoresis and confocal fluorescence microscopy. All grafted nanoparticles were rapidly internalized in HeLa cells. Thus a relationship between the size of th( constructs and their efficacy in nanoparticles delivery was established. A comparison of our resuits with those of the literature shows a promising model for theragnostic devices
Kieleck, Christelle. "Imagerie de fluorescence induite par laser : application à la reconnaissance des groupes de macroalgues." Brest, 2000. http://www.theses.fr/2000BRES2049.
Повний текст джерелаLe, Cardinal de Kernier Isaure. "Cytométrie par imagerie grand champ en phase et fluorescence : applications en hématologie." Thesis, Aix-Marseille, 2019. http://theses.univ-amu.fr.lama.univ-amu.fr/191018_LECARDINALDEKERNIER_341bnro964jhs311fcdqc638rrosk_TH.pdf.
Повний текст джерелаBlood cell population analyses allow detecting a wide scope of clinical disorders, ranging from anemias to malaria. A very large number of cells ought to be considered so as to ensure the statistical significance of the result, and in turn, yield a reliable diagnosis. Currently, hematology analyses are based on flow cytometry techniques. High throughput is obtained at the expense of the information content of each acquisition. To reduce the time-to-result, and to minimize the complexity and cost of the systems dedicated to analyzing cell populations, the current need is to reduce the number of acquisitions and optimize the information content. This thesis focuses on single-shot image cytometry as an alternative to flow-based cytometry. It aims at obtaining a set-up based on optical contrasts for the study of large cell populations while preserving the ability to resolve individual cells. We investigate a multi-scale and multi-modal approach to detect, characterize, and classify blood cells. To evaluate the feasibility and clinical relevance of the method, we developed two proof-of-concept set-ups, respectively called the mesoscope and the miniscope. The mesoscope, based on optical developments, combines phase contrast with fluorescence. The complementarity of morphological features and the expression of specific fluorophores enables us to accurately classify blood cells, and for example assess Plasmodium falciparum parasitemia in whole blood samples. The results are benchmarked to reference techniques. However, to address the need for point of care analyses, the system should be miniaturized. Hence, we designed the miniscope, a chip-based bimodal imager
Pliquett, Jacques. "Development of fluorescent platforms for the design of multifunctional compounds for in vitro and in vivo applications in molecular imaging." Thesis, Bourgogne Franche-Comté, 2018. http://www.theses.fr/2018UBFCK067.
Повний текст джерелаThe objective of this thesis was the development and evaluation of new molecular platformsfor optical fluorescence imaging applications. This work sought to develop new tools that caneasily be modified and adapted to the specific needs of the intended use. This is required asthe fluorophore will influence the final properties and should thus be incorporated beforestructural optimization of the selected agent rather than at the very end. Two main axes wereexplored; the use of BODIPYs for the development of trackable therapeutic agents that areprimarily intended for in vitro applications and the use of azaBODIPYs for the design of an invivo compatible fluorescent platform.In the first part two fluorophores on the basis of a 3,5-dichloro-BODIPY were identified aspromising platforms. These platform molecules were selectively functionalized using a gold(I)-phosphine moiety, a thiosugar and a phosphonium to explore their selective functionalizationand investigate the influence of each substitutents position on the final properties. Weshowed that a site-specific, selective functionalization with these fragile substituents ispossible and developed 12 gold(I)-bearing therapeutic agents. We evaluated thephotophysical properties of all obtained compounds which was followed by a characterizationof their biological properties (antiproliferative properties on 3 cancer cell lines, lipophilicbalance and cellular gold accumulation as well as fluorescence imaging on 3 cell lines for upto 24h). We succeeded in developing a panel of closely related trackable compounds thatdisplay mixed activity in cells and distinct cellular localization. This investigation permitted theselection of three to four hits that will be studied further.In the second part we developed an in vivo-compatible multifunctional platform following twostrategies: the first was the use of 1,7-di(phenol)-3,5-di(phenyl)-azaBODIPY and thefunctionalization of the hydroxy groups for the development of a bioconjugable NIR-I probe.Unfortunately the developed probe displayed very unfavourable optical properties; wetherefore developed a new strategy that is entirely based on the functionalization of the boronatom. Using this approach we successfully synthesized 2 watersoluble, strongly fluorescent(NIR-I) molecular platforms that were conjugated to an innovative antibody to image the PD-L1 ligand. The developed probes displayed excellent optical properties, are stable for at least48h in mice plasma and were validated in a preclinical study on mice. The developed probesdisplayed strong fluorescence in vivo and showed no acute toxicity.The developed methodology shows great potential for further investigations and futurestudies; it can be transposed onto other closely related fluorophores and permits versatilefunctionalization with a large variety of compounds of interest. Its use is thus not limited tobiological, biochemical and medical applications
Nguyen, Anh Dung. "Amélioration de la résolution spatiale en microscopie multiphotonique par saturation de la fluorescence." Doctoral thesis, Universite Libre de Bruxelles, 2015. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/221923.
Повний текст джерела-----------------------Résumé-----------------------Depuis la prédiction de Maria Göppert-Mayer dans les années 30 de la possibilité pour une molécule fluorescente d'être excitée simultanément par plusieurs photons et, plus récemment, depuis le développement des lasers pulsés, la microscopie multiphotonique s'est peu à peu développée pour finalement s'imposer aujourd'hui comme un des outils d'observation par fluorescence les plus performants pour les études de tissus épais diffusants, ou encore pour l'observation in vivo d'animaux. Que ce soit pour des études neurologiques, physiologiques ou morphologiques, l'aspect non invasif et la limitation du volume excité au volume focal ont rendu cet outil de microscopie indispensable aux biologistes.Cependant, dans un monde où les études biologiques nécessitent toujours de meilleurs microscopes et où la résolution spatiale en particulier doit toujours être améliorée, il convient de proposer des techniques permettant d'obtenir une meilleure résolution dans les trois dimensions et d'aller au-delà de la limite de diffraction définie par Ernst Abbe il y a plus d'un siècle.Dans cette thèse, la technique de saturation de l'excitation de la fluorescence est adaptée à la microscopie multiphotonique. Cette méthode permet d'obtenir des images de superrésolution en modulant temporellement l'intensité laser d'excitation et en démodulant les harmoniques supérieures présentes dans le signal saturé de fluorescence. La démonstration de principe sur des microsphères fluorescentes a été réalisée montrant une amélioration de la résolution latérale et axiale. Alors que l'utilisation de la troisième harmonique produit déjà une meilleure résolution, ce travail de thèse montre qu'une amélioration supplémentaire peut être obtenue en utilisant une combinaison linéaire particulière des harmoniques démodulées.Au final, un quasi doublement de la résolution a pu être observé tant dans les directions latérales que dans la direction axiale. Cette amélioration correspond à l'amélioration prédite dans l'analyse théorique et mathématique réalisée également dans ce travail.De plus, le passage aux études in vitro a été réalisé avec succès en observant des microsphères fluorescentes incorporées dans des cellules HeLa. Des améliorations de la résolution latérale et axiale ont également été observées montrant que cette technique de superrésolution peut être appliquée à l'étude d'échantillons biologiques. Les forces et les faiblesses de cette méthode sont également analysées et détaillées afin de voir dans quel créneau d'études biologiques la technique de saturation de l'excitation de fluorescence pourrait se faire une place. A cette fin, ses caractéristiques sont comparées aux autres méthodes de superrésolution et de superlocalisation détaillées dans la première partie de ce travail.Il en resort que l'importante profondeur d'imagerie, l'aspect non invasif et la limitation du volume excité de la microscopie multiphotonique couplés à la simplicité d'implémentation et les relativement faibles puissances utilisées pour saturer l'excitation font de cette technique un excellent candidat pour des études in vivo dans des zones en profondeur dans des milieux diffusants comme la peau.
Doctorat en Sciences de l'ingénieur et technologie
info:eu-repo/semantics/nonPublished
Salomé, Rémi. "Imagerie rapide et suivi de l'activité de réseaux de neurones en microscopie à deux photons." Université Louis Pasteur (Strasbourg) (1971-2008), 2004. http://www.theses.fr/2004STR13063.
Повний текст джерелаLoison, Olivier. "Imagerie plasmonique multimodale en vue d'applications biomédicales." Paris 7, 2013. http://www.theses.fr/2013PA077102.
Повний текст джерелаSurface plasmons are collective oscillation modes of the conductive electrons at the interface between a metal and a dielectric. Plasmonic is a field of optics based on the properties of containment and exaltation of the electromagnetic field given by this mode. Plasmonics is particulary used in biosensors to quantify and to follow interaction dynamic between biomolecules. The specificity of these sensors named Surface Plasmon Resonance (SPR) is their unstrained characteristic. SPR sensitivity relies on the strong dependence of the resonance conditions to the refractive index of the studied dielectric. In particular, the refractive index depends on the biomolecules concentration at the sensor surface. Here, we suggest to use an original device in transmission (tSPR) preserving the strong sensitivity characteristic of standard SPR. We show that this configuration allows a high parallelization density of measurement. Multiplexing given by tSPR is around one order of magnitude more than the one of classical SPR. Propagative surface plasmon (PSP) is associated with an evanescent field. The high intensity of the field and its strong axial confinement have been recently been used in Surface Plasmon mediated Fluorescence (SPF) imaging. Moreover, the presence of a metallic interface in the vicinity of a fluorescent dye induces new relaxation processes, especially via the surface Plasmon, producing a sharp decrease of the fluorescence lifetime. Lifetime information can be extract with a fluorescence lifetime imaging microscope (FLIM). We show that FLIM imaging coupled with surface Plasmon provides nanometric accuracy of the dye-metal distance. We show that such a device is adapted to plasmic membrane tomography with an axial resolution about 15 nm
Montcuquet, Anne-Sophie. "Imagerie spectrale pour l'étude de structures profondes par tomographie optique diffusive de fluorescence." Phd thesis, Université de Grenoble, 2010. http://tel.archives-ouvertes.fr/tel-00557141.
Повний текст джерелаBernardin, Aude. "Méthodes biocompatibles de fonctionnalisation de substrats et surfaces : évaluation par imagerie de fluorescence." Paris 11, 2009. http://www.theses.fr/2009PA112255.
Повний текст джерелаThis work describes the development of biocompatible functionalisation methods of wide interest in the field of biomedical applications. A new water-compatible coupling method between nitrones and strained-cycloalkynes is here developed, by analogy with the [3+2] cycloaddition between azides and strained-cycloalkynes described in the literature. The efficiency of the coupling is evaluated by fluorescence imaging. Two fluorescent probes bearing cyclooctyne groups are synthesized, an organic dye-based probe and a quantum dotbased probe. Contrary to Cu(I)-catalyzed click chemistry, the developped methodology enables quantum dots’ functionalization while maintaining their high fluorescence quantum yield. Glass slides have also been coated with azides or nitrones, surfaces on which can be subsequently grafted cyclooctyne-probes. Fluorescent labelling of cell membrane’s sialic acids after metabolic incorporation of modified mannosamines has also been carried out in vitro to confirm the potential interest of strained-alkyne chemistry in biological conditions
Maréchal, Xavier. "Mise en oeuvre d'une technique de cartographie du pH tissulaire par imagerie fluorescence." Compiègne, 2000. http://www.theses.fr/2000COMP1274.
Повний текст джерелаActually, the mortality rate of septic shock stays important and tissue hypoxia is the major complication of this pathology. Referring literature, measurement of gastro intestinal intramucosal pH (pHim) has been recognized as an important factor in the detection of hypoxia induced dysfonctions. However, current pH measurements techniques are limited in terms of time and spatial resolutions. A major advance in accurate pH measurement was the development of the ratiometric fluorescent indicator dye, 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF). Then, the aim of this study was to setup an in vivo pH measurement technique using BCECF fluorescence imaging. The setup of a new measurement technique is pertormed with two steps: the characterization of the instrumentation and the validation referring a gold standard. These two steps have allowed the setup of the in vivo cartography of pHim using fluorescence imaging and have allowed the validation referring a pH microelectrode. We have then show the feasibility of the intestinal pHim determination and that it is sensitive for in vivo pH determination. Moreover, this fluorescence technique presents a considerable improvement in resolutions compared to current pH measurements techniques. At last, easily transposable to endoscopic measurement, this technique is really interesting for clinical investigations and can be a great advance for early diagnosis of hypoxia induced dysfunction's
Bsaibess, Talia. "Nanoparticules organiques ultra-brillantes pour l'imagerie biologique." Thesis, Bordeaux, 2015. http://www.theses.fr/2015BORD0055/document.
Повний текст джерелаDuring the last decades, luminescent inorganic nanoparticles have attracted a large interest in different fields including biological imaging. However, a number of them have drawbacks such as toxicity and absence of biodegradability. Recently, molecular-based fluorescent organic nanoparticles (FONs) have emerged as a promising alternative to inorganic nanoparticles for bioimaging. The main challenge lies in the elaboration of organic nanoparticles that combine large brightness, good colloidal stability in biological environments) and biocompatibility as well as NIR emission (to allow improved detection in thick tissues). To achieve this objective, we have implemented a molecular engineering strategy based on dedicated polar and polarizable "push pull" chromophore built from a triphenylamine donor moiety and a specific pi-conjugated system. The corresponding nanoparticles were readily prepared by the reprecipitation method. In the present manuscript, the synthesis of the chromophores and the preparation and characterization of the organic fluorescent nanoparticles is described. A comprehensive investigation of their photophysical properties and study of their colloidal stability is presented allowing to derive structure-property relationships. The implemented study led to innovative NIR-emitting nanoparticles combining large brightness (superior to those of QDs and NIR-emitting organic dyes), remarkable colloid stability and suitable photostability. These nanoparticles have been successfully used for single particle tracking and imaging in cells, while no toxic effect was observed
PAGNOT, THIERRY. "Etude temporelle de l'extinction de fluorescence induite par transfert d'energie interloleculaire imagerie en champ proche du temps de declin de fluorescence." Besançon, 1997. http://www.theses.fr/1997BESA2020.
Повний текст джерелаFaucon, Adrien. "Nanomatériaux moléculaires fluorescents et magnétiques pour la bio-imagerie multi-échelle." Nantes, 2014. http://archive.bu.univ-nantes.fr/pollux/show.action?id=19e2fb12-75a7-4527-a030-3b8b6c2acd4a.
Повний текст джерелаThe aim of this PhD work deals with the development, the structural and physico-chemical characterizations of nanomaterials which can combine emission properties after excitation using one or two photons, and magnetism. The association of these two distinct properties, fluorescence and magnetism, allowed us to obtain bimodal functional materials. Over the past years, this kind of nano-objet has raised a large interest in bio-imaging, cancer treatment using hyperthermia, as well as in optical data storage. The bimodal nanomaterials are elaborated by self-assembling iron oxide nanoparticles with small organic fluorophores able to form nanospheres upon nano-precipitation, yielding core-shell nanoassemblies. The development and studies of organic nano-objects, based on small molecules, represent an expanding field, mainly due to the high density of active units. To the best of our knowledge, this tight non-covalent association between a non-doped fluorescent core and a shell composed of iron oxide nanoparticles has never been studied. Before using such magneto-fluorescent nanoassemblies, several studies have to be performed in order to fully understand and characterize the photophysical properties of the fluorophores in solution as well as in the solid state. After a stabilization process compatible with physiological conditions, the resulting nanoassemblies will be used for in vitro and in vivo bioimaging studies. The studies of the biodistribution and the possible degradation of the assemblies will also be performed
Tian, Yayang. "Elaboration of New Layer by Layer (LbL) Fluorescent thin films and their functionalization for the sensitive detection of bacteria." Thesis, Université Paris-Saclay (ComUE), 2018. http://www.theses.fr/2018SACLN029/document.
Повний текст джерелаAntibiotics have been used for the treatment of bacterial infections for over 70 years, saving millions of lives. The current antibiotic resistance crisis has been attributed to the overuse and misuse of these medications. Therefore, the prevention of infection transmission by the rapid and sensitive detection of antibiotic resistant strains is needed in managing this crisis. Fluorescent polymers show great potential for bacteria detection, because they are easy to functionalize, reproduce and graft. Compared with the methods used for bacterial detection in liquid, bacterial detection on a film surface is more convenient, easier to handle and is applied in devices that can be easily reused. The goal of my PhD work is to develop fluorescent and sensitive nanostructured polymer films on surfaces for bacterial detection. Three types of BODIPY-based fluorescent polyelectrolytes (FPC) with different features were synthetized based on reversible addition-fragmentation transfer (RAFT) polymerization: relatively Short chains and Weak polyelectrolytes (SW FPC), Short chains and Strong polyelectrolytes (SS FPCs) and Long chains and Weak polyelectrolytes (LW FPCs). FPC LbL films were fabricated on activated glass slides by means of electrostatic attraction. The photophysical and surface properties of FPC LbL fims were easily controlled by adjusting the deposition conditions.The following step aimed at increasing the films’ sensitivity by using the metal-enhanced fluorescence (MEF) principle. A MEF based LbL film was prepared and tested for bacteria detection. Spherical gold nanoparticles (Au NPs) were synthesized and coated with poly(allylamine hydrochloride) (PAH). The LW FPC- was selected as the fluorescent layer. Different films containing Au NPs and LW FPC- were fabricated and the distance between the Au NPs and LW FPC- was adjusted by changing the numbers of layers with two oppositely charged polymers (PC+ and PC-). Both Au NPs/4 layers PCs/LWFPC- and Au NPs/8 layers PCs/LWFPC- surfaces indicated that E. coli can be detected by LW FPC-.The selectivity of LbL films was added by introducing an antibody on the surface of the film to provide specific recognition of a chosen bacterial strain. This LbL surface achieved a rapid, effective and specific detection of E. coli bacteria. The polyanion and polycation with a 4-dibenzocyclooctynol (DIBO) functional group were assembled on the activated glass slides and an anti-E. coli antibody containing an azide group was efficiently introduced on the surface in a single step based on the azide-alkyne cycloadditions (SPAAC) reaction. The number of E. coli captured on the surface was shown to be dependent on the amount of antibody on the surface. The anti-E. coli antibody surface showed significant selectivity for E. coli, compared with B. subtilis. An alternative approach is to detect bacterial growth on thin LbL film by introducing pH sensitive fluorophore (fluorescein). The growth of bacteria is often associated with a decrease in pH of the growth medium due to a release of acidic metabolites. Different types of pH sensitive LbL film were prepared and tested for the detection of bacterial growth. Firstly, the synthesis of different functionalized polyanions (short and long chain of DIBO-PC- and red fluorescent polymer) was carried out. Three types of pH sensitive surfaces containing fluorescein (DIBO-SWPC-/fluorescein, DIBO-LW PC-/fluorescein and ratiometric RFPC-/fluorescein surfaces) were prepared based on the combination of LbL assembly and copper-free click chemistry. Finally, three pH sensitive surfaces were studied for bacteria growth detection. All the surfaces were shown to be biocompatible, the number of E. coli increased after several hours of incubation on each surface, as detected by brightfield microscopy imaging. The application for the fluorophore-dependent detection of bacterial growth remains to be developed
Hernandez, Mier Yahir Wolf Didier Blondel Walter. "Construction rapide d'images panoramiques applicables à l'exploration cystoscopique et à l'endoscopie de fluorescence en cancérologie." S. l. : INPL, 2007.
Знайти повний текст джерелаDorval, Paul. "Miniaturisation des technologies d'imagerie de fluorescence pour assister la chirurgie mini-invasive." Thesis, Strasbourg, 2015. http://www.theses.fr/2015STRAD005/document.
Повний текст джерелаFluorescence image-guided surgery is a medical imaging modality which allows the surgeon to visualize a fluorescent probe previously injected to the patient. The probe could be specific or not and the technology is useful in a wide range of application from oncologic procedures to reconstructive surgeries or cardiac procedures. Despite the important needs of this technology in open-procedures, the surgery in general is more and more minimally invasive. The goal of mini-invasive surgery is to limit patient's per and post operation trauma. The advantages of the technique are a decrease of bleeding and pain and a decreasing hospitalization time.During an open surgery, the B&W fluorescence information given by the fluorescence image-guided surgery system is enough for the surgeon. For mini-invasive procedures, the in-game is to overlay the fluorescence information to high quality color image, compulsory for the surgeon to perform his procedure. As a first evaluation, a 2-sensors system has been rapidly developed. One sensor is dedicated to the acquisition of the color image and the other to the fluorescence information. In order to make the system more compact and improve the quality of the color image furnished to the surgeon, the final system should be composed of only one sensor. To create the color image and collect the fluorescence information with one sensor, the technique involved pulsed white light and excitation light in a sequential acquisition mode. The two information are combined and a real-time color plus fluorescence video is displayed to the surgeon
Rémond, Maxime. "Développement de nouveaux chromophores dipolaires pour l'imagerie de fluorescence et l'imagerie photoacoustique." Thesis, Lyon, 2018. http://www.theses.fr/2018LYSEN062.
Повний текст джерелаThis thesis is focused on the conception, synthesis and characterization of dipolar dyes D-π-A for fluorescence imaging and photoacoustic imaging.The first part aim to synthesize and to study new fluorophores emitting in the solid-state in the near-infrared for fluorescence imaging with two-photon excitation. In order to improve the optical properties in the solid-state, we first explored various electron donating groups D, then acceptors A and finally we modified the π bridge. In a second part, we focused on dyes for photoacoustic imaging. This technique is based on an optical excitation and an acoustic detection. It requires a strong absorption in the biological window. To red shift the absorption we incorporated a thiophene bridge with low aromaticity to increase the delocalization. We also studied several hemicyanines with strong absorption above 650 nm.Finally, dyes were formulated as water-soluble nanoparticles for their final application for imaging. Two types of nanoparticles were studied: nanoprecipitated dyes stabilized by a surfactant to increase the colloidal stability or encapsulated dyes in a amphiphilic polymer matrix stabilized with a silica shell. Those nanoparticles enabled cells imaging with biphotonic florescence imaging as well as photoacoustic imaging of a microfluidic chip and of the microvascularisation of mice ears in vivo
Pichette, Julien. "Imagerie de fluorescence et intrinsèque de milieux diffusants par temps d’arrivée des premiers photons." Thèse, Université de Sherbrooke, 2014. http://savoirs.usherbrooke.ca/handle/11143/387.
Повний текст джерелаPaganin-Gioanni, Aurélie. "Imagerie de fluorescence d'événements moléculaires et cellulaires associés au cancer au sein d'animaux vivants." Toulouse 3, 2009. http://thesesups.ups-tlse.fr/644/.
Повний текст джерелаFluorescent imaging allows one to monitor spatio-temporal fluorescent events occurring in cells, tissue or living animals and defined in time and space. The development of its use for detecting cancerous cells lies in part in the advancement of probes designed to target and detect. The first axis of my thesis work therefore concerns the evolution of fluorescent probes for the diagnosis of cancer cells in vivo. The second axis will evolve around the study of the transfer of therapeutic molecules in vitro and in vivo to tumoral cells via electropermeabilization. Firstly, the main objective is to develop a "smart molecule" for detecting cancer cells in living organisms by non-invasive fluorescent imaging. Our strategy is based on the use of branched organic molecules called dendrimers, which among their multiple functions allow targeting and detection. In collaboration with Dr. Majoral's team, we attempted to bind several tumour markers to various fluorescent dyes, in order to increase their specificity for B16F10 cancer cells and their signal to noise ratio for detection in vivo. Unfortunately, the chemical nature of dendrimers did not allow us to obtain a functional molecule. Indeed, as they are only soluble in the presence of a high quantity of organic solvent, we encountered a number of problems, due to the denaturing of the biological probes (antibodies) during the binding reaction. In parallel, a variety of non functional dendrimers (cationic, anionic, neutral and fluorescent) were tested on cells in order to determine the most viable candidate for use in vivo. However, we identified problems with the stability of the fluorescence and the toxicity. To solve these issues, with the assistance of a team of chemists, we developed dendrimers bearing polyethylene glycol to enable their dissolution in aqueous environments and make them biocompatible. As these are not cytotoxic, this approach will allow us to elaborate a dendrimer that is functional in an aqueous environment. .
Charonov, Serguei. "Imagerie spectrale en microspectroscopie de diffusion raman et d'emission de fluorescence (doctorat : ingenierie biologique)." Reims, 1999. http://www.theses.fr/1999REIMP207.
Повний текст джерелаJamette, Pascal. "Imagerie de NO par fluorescence induite par laser dans un moteur à allumage commandé." Lille 1, 2001. https://pepite-depot.univ-lille.fr/RESTREINT/Th_Num/2001/50376-2001-273.pdf.
Повний текст джерелаLabouesse, Simon. "Imagerie à éclairements structurés inconnus." Thesis, Aix-Marseille, 2017. http://www.theses.fr/2017AIXM0335.
Повний текст джерелаStructured illumination microscopy (SIM) allow theoretically to double the super-resolution of a standard optical microscope. However, to reach this theoretical limit, SIM require a precise knowledge of the illuminations, making it costly and difficult to calibrate. The aim of this thesis is to simplify the use of SIM by using a blind approach who allow the use of random illuminations to reconstruct a super-resolved image of the object. This strategy theoretically allow the super-resolution, while maintaining a low cost instrumentation. During those three years of thesis, we have studied theoretically and algorithmically the performances and the limitations of a joint estimator of the objet and the illuminations (joint Blind-SIM estimator). A mathematically equivalent reformulation of the joint problem was proposed allowing us to study the super-resolution origin and to propose a fast and parallelizable new approach. An empirical study has highlighted the impact of parsimony and of the frequency content of the illuminations on the reached super-resolution level. Because the joint estimator is asymptotically not consistent, we also studied a contrast criterion for our problem (typically a marginal likelihood), here only the object of interest is estimated. We have mathematically studied the super-resolution capacity of this kind of estimators. Finally, real data using random illuminations where acquired and we have observed a super-resolution effect using our algorithms on multiples real objects of different kind, 2 or 3D, fix or mobile, biological or not, like beads, podosomes, actines
Boucard, Joanna. "Nanomatériaux magnéto-fluorescents pour la bio-imagerie multimodale et la libération contrôlée de principes actifs." Thesis, Nantes, 2018. http://www.theses.fr/2018NANT4051/document.
Повний текст джерелаThese PhD studies consist in the design and characterizations of theranostic nano-tools to meet two requirements. The first one is cell-targeted diagnostics (cancer and muscle cells, bacteria, macrophages) using nanomaterials detectable by fluorescence microscopy and MRI. The second aims at controlling the release of drugs, including anticancerous ones, encapsulated in nanomaterials. The nanometric size of these objects enables enhanced accumulation in tumor tissue (cells), thereby decreasing toxicity side-effects. These objects are supramolecular nanoassemblies composed of a fluorescent solvatochrom organic core coated by a shell of magnetic iron oxide nanoparticles. The emissive core, named FON for fluorescent organic nanoparticle, shows a bright redshifted emission signal compatible with the first biological tissue transparency window. FONs exhibit no cytotoxicity and little photobleaching. The ability of FONs to disassemble once internalized in cellulo and their hydrophobicity enable hydrophobic drugs vectorization. In this way, in cellulo drug delivery can be followed-up by fluorescence microscopy. Drug delivery can be controlled with magnetic hyperthermia thanks to the heating properties due to an alternative magnetic field applied to magnetic nanoparticles. Furthermore, magnetic nanoparticles allow tumor evolution to be monitored in vivo using MRI. All these combined properties pave the way toward theranostic potentialities where personalized nanomedicine is highly requested
Raes, Florian. "Imagerie photoacoustique couplée à l’échographie haute résolution et à la fluorescence infrarouge en oncologie préclinique translationnelle." Thesis, Orléans, 2016. http://www.theses.fr/2016ORLE2082/document.
Повний текст джерелаPreclinical imaging has become an unavoidable step for pathophysiological parameters assessments, for the follow up of tumor growth and for the anticancer therapies development. Technological improvements have emerged in recent years, allowing the emergence of new imaging modalities with a high potential for translation into clinical practice. This manuscript presents several approaches by ultrasound imaging, photoacoustics and near infrared fluorescence in order to monitor the cancer pathology. Initially, we focused on the characterization of hypoxia and its longitudinal assessment in various preclinical models of human cancers. Various multimodal imaging strategies were implemented to assess the efficacy of a new therapeutic prodrug allowing the release of an active molecule in the tumor microenvironment on human models of pancreatic, breast and lung tumors. Finally, in a context of translational research, we explored the potential of photoacoustic and near infrared fluorescence imaging to highlight the lymph node invasion by cancer cells implementing minimally invaded sentinel lymph node models. In this work, we have shown the interest in monitoring the tumor hypoxia in onco-pharmacology and highlighted the high potential of photoacoustic imaging for oncology translational approaches. The main limitation is the relatively shallow depth of regions that we can explore, but this point is currently subject to many technological developments. Feasibility studies performed and validation of proof of concept protocols will enable routine exploitation of these new imaging modalities
Zhang, Ruikang. "Implementations and applications of Out of Phase Imaging after Optical Modulation (OPIOM) in fluorescence macro-imaging and fluorescence endomicroscopy." Thesis, Sorbonne université, 2018. http://www.theses.fr/2018SORUS541.
Повний текст джерелаBased on an analysis of the fluorescent emission on a leaf model, a first optical setup has been built for macro-scale fluorescence imaging, which implements the Speed OPIOM (Out of Phase Imaging after Optical Modulation) protocol to observe the fluorescence of living plants labeled with genetically-encoded reversibly photoswitchable fluorescent proteins. The optical system was designed to image both the red and green emission channels with good quality over an area of about 4 × 4 mm2. This setup allowed biologists to measure the level of expression of fluorescent probes in living plants, without interference from autofluorescence and ambient light, even under sunlight. This fluorescence macroscope also found other applications in fluorescence macro-imaging, e.g. western blot analysis & bacterial selection, against the autofluorescence of the substrate. A fiber bundle-based endoscope incorporating Speed OPIOM has also been built. This second optical setup improved as well the contrast of the image against an autofluorescent background. In addition, it has been used to evidence optical sectioning by a series of numerical simulations and experimental measurements
Khemis, Kamila. "Imagerie de fluorescence en cancérologie : spectroscopie, traitement du signal et gestion automatisée pour l'optimisation du diagnostic des tumeurs précoces." Vandoeuvre-les-Nancy, INPL, 1998. http://docnum.univ-lorraine.fr/public/INPL_T_1998_KHEMIS_K.pdf.
Повний текст джерелаLescure, Robin. "Développement d’azaBODIPYs fonctionnalisables pour la conception de sondes d’imagerie bimodale et d’agents théranostiques." Thesis, Bourgogne Franche-Comté, 2020. https://nuxeo.u-bourgogne.fr/nuxeo/site/esupversions/21e2771a-9c75-46d7-9377-50e8f2536c32.
Повний текст джерелаThe in vivo use of optical imaging is still limited by the lack of near infrared emitting probes. This thesis work focuses on the optimization and valorization of a water-soluble fluorescent platform whose optical properties enable an in vivo use. Two distinct applications were investigated for this WazaBY (Water-soluble azaBODIPY) platform: use as a PET (or SPECT) / optical bimodal probe, and as a theranostic agent. Concerning the first project, we were able to develop a targetted SPECT/optical bimodal probe, which was radiometallated with indium 111. Using xenografted murine models, we were able to show a clear accumulation of the probe in the tumor 24 hours after injection. Moreover, the probe was validated as a contrast agent for fluorescence guided surgery experiment. The second project of this thesis began by the synthesis of a first generation of gold based theranostic agents. The goal was to develop a new therapeutic complex, which can be tracked in vitro and in vivo thanks to optical imaging. In vitro preliminary results showed that the theranostics displayed a cytotoxicity comparable to auranofin on the tested cell lines (4T1, MDA MB 231, CT26 and SW480). A second part of this project focused on the develoment of « smart » probes for a theranostic use. Those probes are designed to undergo photophysical properties changes, when their metallic centre, responsible for the therapeutic role, is released. Two molecules were synthesized, both displaying an on/off behavior
Barilero, Thomas. "Imagerie de la cinétique chimique par modulation de température : conception, validation et mise en oeuvre." Paris 6, 2009. http://www.theses.fr/2009PA066613.
Повний текст джерелаBARANGER, Philippe. "Détection du kérosène par imagerie de fluorescence induite par laser, pour application sur foyer aéronautique." Phd thesis, Université Paris Sud - Paris XI, 2004. http://tel.archives-ouvertes.fr/tel-00009189.
Повний текст джерелаEven, Pascale. "Élaboration de sondes fluorescentes pour des applications en biologie et imagerie par microscopie de fluorescence." Vandoeuvre-les-Nancy, INPL, 2001. http://www.theses.fr/2001INPL567N.
Повний текст джерелаResearchs are done with the aim to develop optical methods for biological studies, with the starting point, the fluorescent trac ers chemical synthesis for two biological applications, namely for biopolymer - cells interactions studies and in photodynamic therapy, respectively. In the first part are described the synthesis and the study of bioactive polysaccharides polymers, labeled with fluorescent molecular rotors. The aim was to synthesise and to study the photophysical properties of coumarin derivatives associated with bioactive dextran (carboxymethyldextranbenzylamide (CMDB)) and heparin. The results of the biological tests prove the interest of the study : fluorescent CMBD polymers keep their stimulation effect on endothelial ceUs growth ; moreover, labeling of endothelial and smooth muscle cells with derivatized fluorescent dextran and heparin respectively, has been demonstrated by fluorescence microscopy. The second part is devoted to the development of new photosensitizers for photodynamic therapy (PDT). PDT is a medical treatment against sorne cancers, based on the combined use of a photosensitizer (often porphyrin compound), oxygen and light. Nowadays, compounds used suffer from sorne drawbacks, especially secondary effects and no specificity. This explains the extend of research in this field, to discover more efficient molecules. Our main objective being the targeting, we developed sorne new porphyrins linked with a variable number of glucosamine rings. Different synthetic routes have been explored and now, the synthese protocols are well known. The photophysical properties of compounds have been studied and aggregate formation, even at low concentration has been observed. Biological tests in course, will orientated the further chemical modifications for a good selectivity
Baranger, Philippe. "Détection du kérozène par imagerie de fluorescence induite par laser, pour application sur foyer aéronautique." Paris 11, 2004. https://tel.archives-ouvertes.fr/tel-00009189.
Повний текст джерелаThe new concepts of aeronautical engines, developed to follow the evolution of the european standards of pollution, are generally based on an improvement of the processes of liquid fuel injection and mixture in the combustion chamber. There is currently no model mature enough to work without experimental validation. The purpose of this thesis is to assess the possibility of measuring the kerosene (jet a1) vapour distribution by plif (planar laser induced fluorescence). That measurement technique must quantitatively image the instantaneous concentrations fields of the vaporized fuel in a spray. The implementation of such a technique needs an experimental spectroscopic study, which was realized on the vapour of fuel. First of all, this study allowed us to determine the properties of the kerosene fluorescence spectrum versus physical parameters such as temperature, pressure or gas mixture composition, especially in presence of oxygen molecules. Then, it was shown that the fluorescence spectrum of the fuel could be reproduce in all physical conditions by a single mixture of four aromatics. Their photophysical properties were also analyzed. Following this spectroscopic study, a phenomenological model for the fluorescence of the gaseous fuel was set up. This model led us to a protocol for an optical diagnostic on this fuel vapour. An experiment was set up to test the implementation and the limits of this technique in simple laboratory conditions. This experiment confirmed that this is indeed a promising technique for the diagnostic of the fuel vapour in aeronautical engine
Detraz, Morgane. "Développement d'outils bimodaux pour l'imagerie oncologique in vivo exploitant les modalités optique et nucléaire." Thesis, Normandie, 2019. http://www.theses.fr/2019NORMR138.
Повний текст джерелаOncologic healthcare and remission prognosis rely on a reliable and accurate diagnosis. Molecular imaging, including optical and nuclear imaging, is currently used for the management cancer therapy. Recently, a new class of bio-imaging agent called MOMIA (MOnomolecular Multimodal Imaging Agent) emerged by combining the synergistic strengths of several modalities on the same molecular structure. We envisioned to merge optical and nuclear imaging in order to develop a molecular tool offering a non-invasive, highly resolutive and sensitive detection. Our approach relies on a universal bimodal clickable scaffold with a selected targeting ligand. Two distinct enzymatic targets have been explored in the oncologic context: caspase-3 as a key component in an apoptotic program and tyrosine kinase inhibitors involved in lung cancer therapy. These multimodal sensors have a promising potential in translational clinical applications
Grichine, Alexeï. "Imagerie spectrale confocale : développement et applications à l'étude des mécanismes d'action photodynamique de nouveaux sensibilisateurs." Orléans, 2002. http://www.theses.fr/2002ORLE2003.
Повний текст джерелаDupont-Therrien, Olivier. "Développement d'outils pour l'imagerie de l'activité neuronale - des épines au comportement." Doctoral thesis, Université Laval, 2017. http://hdl.handle.net/20.500.11794/27491.
Повний текст джерелаDue to the scale of the observed structures and behaviours, neurosciences and microscopy have always been intertwined. Whether it is to observe the different cell morphologies in transmitted white light, or to follow complex dynamics using fluorescent probes, light is the tool of choice to study the brain and its composition. Specifically, the light has the proper spatial and temporal resolution to probe both locally and globally all levels of neuronal activity, while remaining minimally invasive. This thesis shows three techniques developed for different scales, in order to push the limits of the currently addressable biological questions by neuroscientists. In order to effectively probe the internal mechanisms for small structures like dendritic spines and dendrites, we have created a single-cell labeling protocol of the voltage-sensitive fluorophore ANNINE-6plus. The method is based on the intracellular loading of the fluorescent probe in samples both in dissociated cell cultures, than in preparations of acute and organotypic slices. The second project addresses the challenges of rapid imaging of the cellular network activity. Typically, there is a choice between the temporal resolution and the imaging surface. This choice is that the fast imaging techniques are usually widefield and do not provide optical sectioning, making their use in thick samples difficult. By combining a widefield multiphoton technique, the temporal focusing, with the structured illumination and an amplified laser, we have developed a widefield system with an optical sectioning below 10um. The third chapter describes the development of two software distributed with a product created by Doric Lenses Inc., an implantable miniature microscope for imaging of deep brain structures of freely moving animals. This product finally provides a link between the local neuronal activity, and the observed animal behaviours.
Fredj, Asma. "Élaboration de protéines fluorescentes ayant un fort potentiel en imagerie." Phd thesis, Université Paris Sud - Paris XI, 2012. http://tel.archives-ouvertes.fr/tel-00785092.
Повний текст джерелаCanovas, Coline. "Développement de stratégies de bioconjugaison innovantes : Application à l’élaboration d’agents d’imagerie moléculaire." Thesis, Bourgogne Franche-Comté, 2018. http://www.theses.fr/2018UBFCK058.
Повний текст джерелаThe preparation of targeted imaging agents suitable for cancer diagnostic involves the coupling of an imaging probe to a vector, generally a protein, able to specifically target cancerous tissues. Unfortunately, this bioconjugation step is often problematic. Indeed, proteins present a large variety of reactive functions that can potentially interfere with the coupling reaction. The purpose of this work is the design of innovative selective bioconjugation strategies that can facilitate the synthesis of molecular imaging agents.Our research led to the development of a simple and efficient approach allowing the modification of a protein with a fluorophore by using a site-specific bioconjugation reaction. Thanks to this technique, it is possible to obtain near infrared fluorescent agents in a single step.We also focused our work on the design of bimodal nuclear/fluorescent imaging tracers based on protein vectors. These bimodal imaging agents show high potential for numerous medical applications. However, their synthesis requires the coupling of two distinct imaging molecules to the protein, which makes the bioconjugation step even more challenging. In this context, we developed different strategies suitable for the site-specific dual-modification of proteins. These approaches are based on a highly modular trifunctional platform, dichlorotetrazine. Several bimodal nuclear/optical imaging agents were obtained and shown convenient in vivo diagnostic properties.This thesis also presents the elaboration of a new PSMA-specific radiotracer able to detect prostate cancer. The promising results obtained during the preclinical evaluation of this compound labeled with copper-64, make it an interesting candidate for the diagnosis of prostate cancer by positron emission tomography in human
Gavoille, André. "Application de l'analyse d'images à une classe d'images biologiques particulière : les images de tâches." Paris 12, 1989. http://www.theses.fr/1989PA120047.
Повний текст джерелаConrath, John. "Caractérisation de la zone avasculaire centrale chez le diabétique en angiofluorographie rétinienne : corrélations quantitatives et qualitatives, influence des facteurs métaboliques et essai de détection semi-automatisée." Aix-Marseille 2, 2004. http://www.theses.fr/2004AIX20671.
Повний текст джерелаDiabetic retinopathy is the second cause of legal blindness in France and the first cause in the less than 50 years old age group. Retinopathy is one of the three manifestations of diabetic microangiopathy along with nephropathy and neuropathy. Retinal ischemia in diabetes affects peripheral retinal vessels as well as macular vessels. Peripheral ischemia is well known and an effective treatment exists, panretinal photocoagulation. Macular ischemia has been less studied perhaps because there exists no effective treatment for it. The center of the retina is physiologically avascular. This Foveal Avascular Zone enlarges during diabetes. We used our digital image bank of the University Hospital Center of Marseille's ophthalmology departments in a retrospective study of 110 diabetic patients to caracterise the FAZ. We found a correlation between quantitative and qualitative analysis of the FAZ (using the ETDRS grading system) (p=0. 03), with the FAZ enlarging as the grade of macular vessel destruction increased. We found a significantly higher body mass index in patients presenting a normal FAZ (p=0. 033) perhaps because certain type 2 diabetics not requiring insulin present a less severe form of the disease. We also found a protective effect of the Apo A1 fraction of serum cholesterol on the FAZ in the sub-group having undergone that measure (p=0. 004). The FAZ may be measured with an image analysis software programme (ENVI), but it strongly depends upon image quality, and underestimates FAZ surface compared to manual measurement
Nouizi, Farouk. "Tomographie optique diffuse et de fluorescence préclinique : instrumentation sans contact, modélisation et reconstruction 3D résolue en temps." Phd thesis, Université Louis Pasteur - Strasbourg I, 2011. http://tel.archives-ouvertes.fr/tel-00658285.
Повний текст джерелаChomik, Alain. "Déconvolution 3D orientée vers la reconstruction d'objets biologiques observés en microscopie optique de fluorescence." Mulhouse, 1997. http://www.theses.fr/1997MULH0470.
Повний текст джерелаManandhar, Sandeep. "3D motion estimation and assessment in fluorescence microscopy volume sequences." Thesis, Rennes 1, 2019. http://www.theses.fr/2019REN1S096.
Повний текст джерелаThe thesis work deals with the computation and the assessment of 3D motion fields in 3D fluorescence microscopy image sequences. We have investigated 3D matching and variational methods for 3D flow field estimation between two consecutive volumes. For matching, we have developed two original 3D extensions of PatchMatch both involving the discrete Census similarity measure: a super-pixel based method that proceeds slice by slice, and a coarse-to-fine method directly applied to the volumes. We have also designed a protrusion segmentation method on the cell surface along with a matching stage relying on a triangular mesh-based representation. Regarding the dense estimation of 3D flow fields, we have adopted a variational approach, while exploiting the continuous Census signature of voxels in the data term. We have tested three regularization terms: L2, L1, and TV-based regularization. We have also combined the 3D PatchMatch method with the variational method to be able to handle simultaneously large and small motion magnitude. For visual assessment, we have proposed three different color-coded visualization techniques of 3D flow fields. They offer 2D summaries of the 3D flow field, respectively, slice-by-slice, with tri-planar projections, and after maximum intensity point projection. In addition, we have defined a new quantitative error measure for assessing the accuracy of the estimated flow field when no ground truth is available. It involves the angular difference between the local principal orientation of the original source volume and the corresponding one in the volume backward-warped with the 3D computed flow field. We have tested our methods on real microscopy image sequences containing MV3 melanoma cells in collagen environment. When comparing with the state-of-the-art method of Amat et al., and our 3D extension of the classical Horn-and-Schunck method, we found our proposed methods to be the best performing ones
Sivankutty, Siddharth. "Imaging beyond the diffraction limit STED and SAF microscopy." Thesis, Paris 11, 2014. http://www.theses.fr/2014PA112108.
Повний текст джерелаUnderstanding cellular processes on membranes has been a key area of biomedical research. Circumventing the diffraction limit in fluorescence microscopy has now become possible by exploiting the molecular transitions of the fluorophore. In this context, this work presents the instrumental development of two complementary techniques for realizing nanometric all-optical resolution and axial sectioning, namely STimulated Emission Depletion (STED) and Supercritical Angle Fluorescence (SAF) microscopy. STED microscopy is an elegant method that has allowed us to break the diffraction barrier with light microscopes and has achieved resolutions of the order of 40 nm (transverse) in biological samples. In this technique, we exploit the molecular transitions of the fluorescent marker to overcome the resolution limit due to diffraction. Resolution enhancement is achieved by efficient depletion of the excited state of the marker in the peripheral spatial regions of the focal volume by using depletion beams in addition to the excitation beam. Despite the major resolution improvement demonstrated, the technique is not well spread out, mainly due to its apparent complexity; and the cost and limited tunability of the commercial system. In this context, the instrumental realization and the imaging performance of a cost-effective home-built STED microscope is presented in this manuscript. While conventional STED microscopes offer improved lateral resolution, an isotropic gain in resolution usually comes at the cost of complex instrumentation. In this regard, we demonstrate SAF microscopy as a powerful tool that achieves an axial sectioning of the order of 150 nm. This is done by exploiting the property of a molecule to emit into the supercritical anglesonly when near the glass-water interface. Axial sectioning is obtained in a simple configuration by detecting solely the supercritical components of radiation. A combination of these imaging techniques offer a powerful tool to study molecular phenomena on the biological membranes
Maurice, Vincent. "Fonctionnalisation de nanoparticules de silicium pour l’imagerie biomédicale." Paris 11, 2010. http://www.theses.fr/2010PA112355.
Повний текст джерелаWe studied and modified the properties of silicon nanoparticles produced by laser pyrolysis, to use them as fluorescent probes for biological imaging. At first, the surface properties of the particles have been studied, especially their oxidation mechanisms. The silicon particles are indeed easily oxidized in aqueous medium, leading to their destruction if this phenomenon is not controlled. Knowledge of factors causing the deterioration of the properties of particles in biological media has allowed us to define the functionalization strategy adopted for this study. We chose to protect the particles with a layer of silica formed by sol-gel process in homogeneous phase or microemulsion, its thickness ranging from 1 to 30 nm. The encapsulated particles have been characterized, in particular, their rate of oxidation at neutral ph was measured. They were then covered with amines in order to graft organic molecules to their surface. PEG chains were grafted onto the amines to improve the colloidal stability of the particles and their resistance to oxidation. Then we performed the grafting of a protein, to provide biological functionality to the particles. The various functions grafted were assayed by different methods. The particles were also used for biological imaging tests as well as measures of toxicity
Tramier, Marc. "Imagerie des déclins de fluorescence pour l'étude de la dynamique et des interactions de macromolécules en cellules vivantes." Phd thesis, Université Pierre et Marie Curie - Paris VI, 2001. http://tel.archives-ouvertes.fr/tel-00003477.
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