Molecular imaging plays an important role for development of systems biomedicine, which non-invasively extracts pictorial information on physiological and pathological activities at the cellular and molecular levels. Optical molecular tomography is an emerging area of molecular imaging. It locates and quantifies a 3D molecular probe distribution in vivo from data measured on the external surface of a small animal around the visible and infrared range. This approach can facilitate or enable preclinical applications such as cancer studies, involving angiogenesis, tumor growth, cell motility, metastasis, and interaction with a micro-environment. The reconstruction of diffuse light sources is the central task of optical molecular tomography, and generally ill-posed and rather complex. The key element of optical molecular tomography includes the geometrical model, tissue properties, photon characteristics, transport model, and reconstruction algorithm. This dissertation focuses mainly on the development optical molecular tomography methods based on bioluminescence/fluorescence probes to solve some well-known challenges in this field. Our main results are as follows. We developed a new algorithm for estimation of optical parameters based on the phase-approximation model.  Our iterative algorithm takes advantage of both the global search ability of the differential evolution algorithm and the efficiency of the conjugate gradient method. We published the first paper on multispectral bioluminescence tomography (BLT). The multispectral BLT approach improves the accuracy and stability of the BLT reconstruction even if data are highly noisy. We established a well-posed inverse source model for optical molecular tomography. Based on this model, we proposed a differential evolution-based reconstruction algorithm to determine the source locations and strengths accurately and reliably. Furthermore, to enhance the spatial resolution of fluorescence molecular tomography, we proposed fluorescence micro-tomography to image cells in a tissue scaffold based on Monte Carlo simulation on a massive parallel processing architecture. Each of these methods shows better performance in numerical simulation, phantom experiments, and mouse studies than the conventional methods. / Ph. D.

L’imagerie moléculaire optique joue maintenant un rôle essentiel dans le diagnostic pré-clinique et le développement de médicaments. En effet, c’est un outil précieux dans la détection et le suivi de cellules vivantes que ce soit en utilisant de simples agents de marquage ou des sondes plus développées, dites « intelligentes » et activées uniquement par une interaction spécifique avec le bio-analyte ciblé. Ce travail de thèse a consisté à développer des outils synthétiques innovants afin d’optimiser les paramètres physico-chimiques et les propriétés optiques des sondes luminescentes. Ceci dans le but de répondre à la problématique complexe de l’imagerie dans le contexte in vivo. Nous avons notamment travaillé sur des aspects de pro-fluorescence et de chémiluminescence. De nouveaux pro-fluorophores à phénol basés sur une architecture originale de type bis-coumarinique ont été développés. De plus, nous avons mis en place une méthode d’hydrosolubilisation généralisable aux fluorophores à phénol de type coumarine et xanthène. Nos recherches en chémiluminescence ont permis la synthèse de nouveaux chémiluminophores couplés à des fluorophores organiques afin d‘augmenter l’efficacité d’émission de chémiluminescence dans le rouge. Enfin, nos travaux ont permis de mettre en place les premières « cassettes » chémiluminescentes basées sur une architecture de type 1,2-dioxétane. / Optical molecular imaging is now playing a pivotal role both in pre-clinical diagnosis and drug development. Indeed, this is a valuable tool for the real time detection and monitoring of living cells either through the use of structurally simple labels or more recently by means of sophisticated fluorescent probes, called “smart” probes and only activatable upon specific interaction with the targeted bio-analyte. The aim of this PhD work was the design of new synthetic tools aimed at optimizing physico-chemical and optical properties of fluorescent probes intended for challenging in vivo imaging applications. We have focused on the pro-fluorescence and chemiluminescence approaches. New phenol-based pro-fluorophores have been developed by using an original bis-coumarinic scaffold. In the context of the chemistry of fluorophores, we have also investigated a general method for the water-solubilisation of phenol-based fluorophore belonging to the coumarin and xanthene families. Our research in chemiluminescence has led the synthesis of new chemiluminophores covalently linked to fluorescent organic dyes aimed at increasing the emission efficiency in the red region of such chemiluminophores. Thus, the first chemiluminescent “energy transfer cassettes” based on a 1,2-dioxetane scaffold have been obtained.