No description available.

Optical coherence imaging, including tomography (OCT) and microscopy (OCM), has been a growing research field in biomedical optical imaging in the last decade. In this imaging modality, a broadband light source, thus of short temporal coherence length, is used to perform imaging via interferometry. A challenge in optical coherence imaging, as in any imaging system towards biomedical diagnosis, is the quantification of image quality and optimization of the system components, both a primary focus of this research. We concentrated our efforts on the optimization of the imaging system from two main standpoints: axial point spread function (PSF) and practical steps towards compact low-cost solutions. Up to recently, the criteria for the quality of a system was based on speed of imaging, sensitivity, and particularly axial resolution estimated solely from the full-width at half-maximum (FWHM) of the axial PSF with the common practice of assuming a Gaussian source power spectrum. As part of our work to quantify axial resolution we first brought forth two more metrics unlike FWHM, which accounted for side lobes in the axial PSF caused by irregularities in the shape of the source power spectrum, such as spectral dips. Subsequently, we presented a method where the axial PSF was significantly optimized by suppressing the side lobes occurring because of the irregular shape of the source power spectrum. The optimization was performed through optically shaping the source power spectrum via a programmable spectral shaper, which consequentially led to suppression of spurious structures in the images of a layered specimen. The superiority of the demonstrated approach was in performing reshaping before imaging, thus eliminating the need for post-data acquisition digital signal processing. Importantly, towards the optimization and objective image quality assessment in optical coherence imaging, the impact of source spectral shaping was further analyzed in a task-based assessment method based on statistical decision theory. Two classification tasks, a signal-detection task and a resolution task, were investigated. Results showed that reshaping the source power spectrum was a benefit essentially to the resolution task, as opposed to both the detection and resolution tasks, and the importance of the specimen local variations in index of refraction on the resolution task was demonstrated. Finally, towards the optimization of OCT and OCM for use in clinical settings, we analyzed the detection electronics stage, which is a crucial component of the system that is designed to capture extremely weak interferometric signals in biomedical and biological imaging applications. We designed and tested detection electronics to achieve a compact and low-cost solution for portable imaging units and demonstrated that the design provided an equivalent performance to the commercial lock-in amplifier considering the system sensitivity obtained with both detection schemes.

L’étude du cerveau humain est un domaine en plein essor et les techniques non-invasives de l’étudier sont très prometteuses. Afin de l’étudier de manière non-invasive, notre laboratoire utilise principalement l’imagerie par résonance magnétique fonctionnelle (IRMf) et l’imagerie optique diffuse (IOD) continue pour mesurer et localiser l’activité cérébrale induite par une tâche visuelle, cognitive ou motrice. Le signal de ces deux techniques repose, entre autres, sur les concentrations d’hémoglobine cérébrale à cause du couplage qui existe entre l’activité neuronale et le flux sanguin local dans le cerveau. Pour être en mesure de comparer les deux signaux (et éventuellement calibrer le signal d’IRMf par l’IOD), où chaque signal est relatif à son propre niveau de base physiologique inconnu, une nouvelle technique ayant la capacité de mesurer le niveau de base physiologique est nécessaire. Cette nouvelle technique est l’IOD résolue temporellement qui permet d’estimer les concentrations d’hémoglobine cérébrale. Ce nouveau système permet donc de quantifier le niveau de base physiologique en termes de concentrations d’hémoglobine cérébrale absolue. L’objectif général de ma maîtrise était de développer un tel système afin de l’utiliser dans une large étude portant sur la condition cardiovasculaire, le vieillissement, la neuroimagerie ainsi que les performances cognitives. Il a fallu tout d’abord construire le système, le caractériser puis valider les résultats avant de pouvoir l’utiliser sur les sujets de recherche. La validation s’est premièrement réalisée sur des fantômes homogènes ainsi qu’hétérogènes (deux couches) qui ont été développés. La validation des concentrations d’hémoglobine cérébrale a été réalisée via une tâche cognitive et appuyée par les tests sanguins des sujets de recherche. Finalement, on présente les résultats obtenus dans une large étude employant le système d’IOD résolue temporellement en se concentrant sur les différences reliées au vieillissement. / Our understanding of the functional organization of the human brain has been greatly influenced by the development of new medical imaging techniques. Pr. Hoge’s research has focused on the use of functional magnetic resonance imaging (fMRI) and continuous diffuse optical imaging (DOI) for non-invasive localization and quantification of brain activity associated with behavioral stimuli or tasks (e.g. cognitive, motor or visual). The respective signals of both techniques are based on cerebral haemoglobin concentrations because of the coupling that exists between neuronal activity and cerebral blood flow. Relating BOLD fMRI signals with those acquired using DOI has been complicated by the fact that fMRI yields fractional change values, while the majority of DOI methods have provided absolute changes from an unknown baseline. To address this, we adopted a newer technique known as time-resolved DOI, which allows absolute quantification of cerebral haemoglobin concentrations. Time-resolved DOI thus has the capacity to quantify the subject’s resting hemoglobin concentrations in absolute micromolar units. The main objective of my masters’ project was to implement and optimize a time-resolved DOI system for use in a large study exploring the links between cardiovascular fitness, aging, neuroimaging markers, and cognitive performance. In this thesis we describe the fabrication of the system, followed by its characterisation and validation using solid optical phantoms (homogeneous and heterogeneous) developed for this purpose. Haemoglobin concentrations obtained non-invasively with the system are validated against blood draws, while the sensitivity to variations in concentration are assessed during a cognitive task. Finally, we present the results of a large study in which the time-resolved DOI system was used to characterize age-related vascular changes in the brain.

L’optogénétique est une technique prometteuse pour la modulation de l’activité neuronale. Par l’insertion d’une opsine microbienne dans la membrane plasmique de neurones et par son activation photonique, il devient possible de réguler l’activité neuronale avec une grande résolution temporelle et spatiale. Beaucoup de travaux ont été faits pour caractériser et synthétiser de nouvelles opsines. Ainsi, plusieurs variétés d’opsines sont désormais disponibles, chacune présentant des cinétiques et sensibilités à des longueurs d’onde différentes. En effet, il existe des constructions optogénétiques permettant de moduler à la hausse ou à la baisse l’activité neuronale, telles la channelrhodopsine-2 (ChR2) ou la halorhodopsine (NpHR), respectivement. Les promesses de cette technologie incluent le potentiel de stimuler une région restreinte du cerveau, et ce, de façon réversible. Toutefois, peu d’applications en ce sens ont été réalisées, cette technique étant limitée par l’absorption et la diffusion de la lumière dans les tissus. Ce mémoire présente la conception d’une fibre optique illuminant à un angle de 90° à sa sortie, capable de guider la lumière à des structures bien précises dans le système nerveux central. Nous avons conduit des tests in vivo dans le système visuel de souris transgéniques exprimant la ChR2 dans l’ensemble du système nerveux central. Dans le système visuel, les signaux rétiniens sont conduits au corps genouillé latéral (CGL) avant d’être relayés au cortex visuel primaire (V1). Pour valider la capacité de mon montage optogénétique à stimuler spécifiquement une sous-population de neurones, nous avons tiré profit de l’organisation rétinotopique existant dans le système visuel. En stimulant optogénétiquement le CGL et en tournant la fibre optique sur elle-même à l’aide d’un moteur, il devient possible de stimuler séquentiellement différentes portions de cette structure thalamique et conséquemment, différentes représentations du champ visuel. L’activation des projections thalamiques sera enregistrée au niveau de l’aire V1 à l’aide de l’imagerie optique intrinsèque, une technique qui permet d’imager les variations de la concentration d’oxygène et du volume sanguin dans le tissu neuronal, sur une grande surface corticale. Comme l’organisation rétinotopique est maintenue au niveau de l’aire V1, l’espace activé au niveau du cortex révèlera l’étendue spatiale de notre stimulation optogénétique du CGL. Les expériences in vivo démontrèrent qu’en déplaçant la fibre optique dans le CGL, il nous était possible de stimuler différents sous- ensembles de neurones dans cette structure thalamique. En conclusion, cette étude montre notre capacité à développer un système à base de fibre optique capable de stimuler optogénétiquement une population de neurone avec une grande précision spatiale. / Optogentics is a promising technic for neuronal activity modulation. By inserting a microbial opsin in the plasma membrane and by its photonic activation, it is possible to regulate neuronal activity with high temporal and spatial resolution. A lot of work has been done to characterize and synthetize new opsins. Thus, a wide variety of opsins are now available, presenting different kinetics and sensibility to specific wavelengths. Indeed, different opsins can either increase or decrease neuronal activity such as channelrhodopsin-2 (ChR2) or halorhodopsin (NpHR), respectively. This technology has the potential to stimulate a specific region within the brain in a highly reversible manner. However, little work was accomplished in this way, because to limitations due to absorption and scattering of light in biological tissue. This master’s thesis presents the conception of a side-firing optical fiber, capable of guiding light to specific structures within the brain. We conducted in vivo experiments in the visual system of transgenic mice expressing ChR2 in the entire central nervous system. In the visual system, retinal inputs are relayed to the lateral geniculate nucleus (LGN) before reaching the primary visual cortex (V1). To validate the capacity of the designed optogenetic assembly to stimulate specific sub-populations of neurons, we took advantage of the retinotopic organization existing in the visual system. By optogenetically stimulating the LGN and rotating the optical fiber around its axis with a motor, it is possible to sequentially stimulate different portions of this thalamic structure and consequently, different portions of the visual field. Activation of thalamic projections will be recorded in area V1 using intrinsic optical imaging, a technic allowing to image variations of blood oxygenation and blood volume in neuronal tissue over large cortical areas. Activation at the level of the cortex will reveal the spatial extent of the optogenetic stimulation in the LGN as retinotopic organization is maintained in V1 cortical area. In vivo experiments showed that displacing the optical fiber in the LGN allowed stimulation of different neuronal populations within this thalamic structure. In conclusion, this study demonstrates our capacity to develop a fiber-based system capable of optogenetically stimulating neuronal tissue with high spatial precision.

Malgré le rôle primordial des neurotransmetteurs (NTs) dans le système nerveux central, leur détection non-invasive in vivo reste un défi majeur. L’imagerie par résonance magnétique (IRM), grâce à son excellente résolution spatiale et temporelle, est parmi les techniques de diagnostic les plus performantes. Elle est au centre des développements récents en imagerie moléculaire. En particulier, l’utilisation des agents d’imagerie intelligents qui sont capables de visualiser le statut physico-chimique des tissus commence à avoir une place importante en neuroscience.Cette étude a pour objectif de concevoir, synthétiser et caractériser in vitro des sondes intelligentes à base de cations lanthanide pour la détection in vivo des NTs. La conception de nos sondes est basée sur des interactions doubles avec des neurotransmetteurs zwitterioniques: d’une part entre le complexes de Ln3+ positivement chargé et le carboxylate du NT et d’autre part entre un ether couronne lié au complexe et la fonction amine du NT. Plusieurs des sondes synthétisées présentent des relaxivités élevées et ont une réponse relaxometrique remarquable aux NTs, bien que leur sélectivité vis-à-vis de l’ion bicarbonate ne soit pas suffisante. Afin de développer des sondes pour une approche bimodale IRM /optique, nous avons également intégré dans les complexes une benzophenone qui joue le rôle de chromophore pour sensibiliser la luminescence des ions Ln3+ émettant dans le proche infra-rouge. Le complexe d’Yb3+ correspondant a des propriétés de luminescence très intéressantes avec une forte réponse aux NTs. / In spite of the key role of neurotransmitters (NTs) in signal transduction, their non-invasive in vivo monitoring remains an important challenge. Magnetic resonance imaging (MRI) has recently been demonstrated as a promising technique to non-invasively visualize physiological events with excellent temporal and spatial resolution. In particular, smart MRI contrast agents that are able to report on the physico-chemical status of the tissues, start to have a strong impact in neuroscience. The objective of this work was the design, synthesis and in vitro characterization of a series of lanthanide-based probes responsive to NTs with the aim to track in vivo concentration changes of NTs using MR or optical imaging. The design of our imaging probes relies on a dual binding approach of zwitterionic NTs to the Ln3+ complexes, involving interactions (i) between a positively charged Ln3+ chelate and the carboxylate function of the NTs and (ii) between an azacrown ether appended on the chelate and the amine group of the neurotransmitters. Some of the novel contrast agents were found to exhibit high relaxivities and a remarkable relaxivity response towards NTs, though little selectivity against bicarbonate. In order to apply a bimodal MRI/optical imaging approach, we have also incorporated a benzophenone moiety into the chelate to sensitize the near-infrared emitting Ln3+ ions. The Yb3+ analogue proved to be highly sensitive to NTs.

Nous avons analysé l'activité de population au sein du cortex visuel primaire en vue de comprendre (i) les mécanismes mis en jeu lors de l'intégration de l'information visuelle suite à un mouvement oculaire, et inversement (ii) de l'influence du traitement effectué au niveau de V1 sur la génération d'un mouvement oculaire.1. Nous avons enregistré des saccades de fixation, et mis en relation, essai par essai, ces mouvements avec la représentation de la position d'un stimulus local dans V1. Après une saccade de fixation, l'activité se déplace de façon cohérente dans V1. Le décours temporel des réponses au niveau des foyers pre- et post-saccadiques montre une dynamique biphasique. La taille du foyer d'activité augmente. Nous proposons que le comportement des populations de neurones s'explique par deux phénomènes principaux : (i) La réponse suppressive précoce attribuable à la décharge corollaire (ii) de connections latérales qui réactiveraient le foyer pre-saccadique.2. Nous avons enregistré l'OFR, et cherché à savoir si la réponse de V1 l'influençait. Les latences VSD précèdent les latences OFR. Il n'existe pas de corrélation à l'essai unique. Nous avons montré que la force et la dynamique des réponses de V1 n'étaient pas prédictives de l'OFR. La distance de la périphérie à un effet sur la réponse VSD, mais pas sur l'OFR. La dynamique de propagation de cette suppression, nous avons montré deux phases : une précoce sur l'ensemble de la carte, et une plus périphérique tardive. Nous proposons que la suppression précoce soit originaire de projections en retour de structures comme MT et MST, alors que la suppression plus lente s'explique par les connections horizontales. / We analyzed population activity in V1 to understand (i) the consequence of eye movements on integration of visual information, and (ii) the influence of the processing performed at the level of V1 on the generation of eye movements.1. We recorded fixational saccades, relating, trial-by-trial, these eye movements with the representation of the position of a local stimulus in V1. After a fixational saccade, activity moves consistently in V1. However, the time-course of responses display a biphasic dynamic. This results in a global increase of the extent of cortical activity representing the local stimulus. We propose that the behavior of populations of neurons studied is explained by the contribution of two main phenomena: (i) an early suppressive response that could be attributed to the corollary discharge and (ii) the lateral connections generating lateral interactions between pre and post-saccadic lci of activity.2. We recorded the ocular following response, determining whether the response of V1 influences the oculomotor response. We studied the contrast response function of the population V1 activity and the OFR. The dynamics of CRF for a local stimulus are similar and shifted in time. We found no correlations between the single trial latencies between V1 and the OFR. At the chosen scale, surround suppression was found to be distance-dependent only in V1. The dynamics of the surround suppression shows two phases: an early suppression present over a wide cortical area, and a later peripheral spread. We propose that the early surround suppression originates from feedback from MT and MST, while the later is explained by the horizontal connections.

Vasoaktives Intestinales Peptid (VIP) ist ein 28meres Neuropeptid, das eine Vielzahl biologischer Aktivitäten ausübt, welche durch die Bindung an die heptahelikalen Transmembranrezeptoren VPAC1 und VPAC2 vermittelt werden. VPAC1 ist auf der Oberfläche vieler Tumorzellen überexprimiert. Daher ist VIP gekoppelt mit Signalmolekülen wie z.B. Fluoreszenzfarbstoffen eine interessante Zielstruktur für die optische Detektion von Tumoren. Der Einsatz von VIP in der Tumordiagnostik ist jedoch aufgrund der schnellen proteolytischen Degradation stark limitiert. In der vorliegenden Arbeit wurden VIP-Analoga mit höherer in vitro und in vivo Stabilität identifiziert und synthetisiert. Hierfür wurde eine neue Synthesestrategie entwickelt, welche die hochparallele Herstellung von löslichen VIP-Farbstoff-Konjugaten auf Cellulosemembranen (Spotsynthese) erlaubt. Es wurden 533 N-terminal Carbocyanin-farbstoff-markierte VIP-Analoga synthetisiert, wobei jeder VIP-Rest durch alle übrigen 19 L-Aminosäuren ausgetauscht wurde (Substitutionsanalyse). Alle Analoga wurden mittels Durchflußzytometrie hinsichtlich ihrer Bindung/Internalisierung an VPAC1-überexprimieren-den Zellen getestet. Diese Ergebnisse führten zur Identifizierung von VIP Aminosäureresten, die für die Wechselwirkung mit VPAC1 essentiell sind und lieferten weiterhin Hinweise über eine vorwiegend helikale Struktur des Rezeptor-gebundenen VIPs. Durch Einbau des Farbstoffs an alle VIP-Reste wurden die für die Wechselwirkung mit VPAC1 günstigen Positionen ermittelt. In Kompetitionstudien und cAMP-Assays ausgewählter Farbstoff-markierter VIP-Konjugate wurde demonstriert, daß sowohl die Spezifität als auch die Produktion von cAMP mit Hilfe der Modifikation der Farbstoffposition gesteigert werden konnte. Für die in Rattenleber getestete metabolische Stabilität der VIP-Analoga zeigte die Farbstoffposition nur einen geringen Einfluß. Die metabolische Stabilität konnte jedoch durch eine einzige Modifikation an Position 8 (Asp8 ® Arg8) erhöht werden. Weiterhin wurde das [Arg8]-VIP-Analogon als Kontrastmittel in in vivo Imaging-Experimenten mit VPAC1-überexprimierenden Tumoren inokulierter Mäuse appliziert. Hierbei wurden die Ergebnisse der Stabilitätstests in Rattenleber bestätigt. Das N-terminal farbstoffmarkierte [Arg8]-VIP-Derivat zeigte gegenüber dem nativen N-terminal markierten VIP-Konjugat eine höhere Halbwertszeit in vivo. Darüber hinaus konnte mit [Arg8]-VIP ein höherer Fluoreszenzkontrast zwischen normalen und Tumorgewebe induziert werden. / Vasoactive Intestinal Peptide (VIP) is a 28meric neuropeptide with a broad range of biological activities which are mediated via binding to the heptahelical transmembrane receptors VPAC1 and VPAC2. Since VPAC1 is overexpressed on the surface of numerous tumour cells, VIP coupled with signal structures like fluorescence dyes is an interesting target for the tumour detection in the near-infrared range. The use of VIP in tumour diagnosis is limited due to the rapid proteolytic degradation and therefore suggests need for optimised VIP-analogues with enhanced stability. In the present work a complete substitutional analysis of VIP N-terminally labelled with carbocyanine dyes was performed. For that reason a new synthetic strategy has been developed, which allows the parallel production of soluble VIP-dye conjugates using the spot synthesis technique. The resulting 560 derivatives were tested for binding and internalisation using VPAC1-overexpressing cells by means of flow cytometry. Based on these results a VIP binding motif has been delineated, which facilitates the modification of VIP concerning stability enhancement and preservation of binding characteristics. Using a dye-walk the dye-coupling positions beneficial for cell binding were identified. Radioactive competitions studies and cAMP assays of selected dye-labeled VIP-conjugates demonstrated that specifity as well as production of cAMP or biological activity has been increased by alteration of the dye-position. For increasing metabolic stability of labelled VIP-analogues the dye-position showed little influence. But is has been shown that stability of VIP was increased by only one modification at position 8 (Asp8®Arg8). Furthermore [Arg8]-VIP-derivative has been used as a contrasting agent in in vivo-Imaging experiments with VPAC1-overexpressing tumours inoculated in mice. Here the results of stability test in rat liver extract were confirmed. N-terminally dye-labelled [Arg8]-VIP analogue revealed higher half-life-time in vivo towards N-terminally labelled native VIP-derivative. In addition the [Arg8]-VIP induced a higher tumour contrast between normal and tumour tissue.

Au cours des dernières années, l’imagerie médicale est devenue l’une des techniques les plus puissantes dans le domaine du diagnostic médical et des recherches biomédicales. Avec le développement de l’imagerie moléculaire, les sondes sensibles permettant l’imagerie multimodale des événements moléculaires sont alors nécessaires.Dans ce travail, nous présentons la conception et la synthèse des complexes de lanthanides dans le but de développer des agents de contraste intelligents pour la détection de l’activité enzymatique par IRM (T1/CEST) et l’imagerie optique. Les complexes conçus s’articulent autour d’un chélate de lanthanide macrocyclique joint avec une amino pyridine qui est liée à un déclencheur enzymatique sensible (e.g. galactoside) par un espaceur auto-immolatif. Celui-ci est censé modifier temporairement, en fonction de la présence d’enzyme, les propriétés magnétiques et photo-physiques du complexe. Le concept a été validé sur un composé modèle sans déclencheur. Bien qu’aucune différence de relaxation n’ait été observée entre les modèles de forme enzymatique activée et non-activée qui empêche l’utilisation de T1-IRM, des effets ParaCEST différents dépendant du lanthanide, ont été observé. En outre, un effet CEST inconnu a été affecté à la fonction carbamate. Des études photo-physiques préliminaires ont montré également des propriétés différentes des deux formes et ont confirmé le potentiel de ces complexes comme agents de contraste enzymatiques sensibles bimodals. La synthèse de la sonde enzymatique sensible a été tenté par trois voies différentes et a finalement été effectué dans un processus de treize étape qui restait à être optimisé. Une étude sur la relation ‘structure-activité’ a été lancée avec la synthèse des isomères de position sur la pyridine du composé modèle / Over the last decade, medical imaging has evolved into one of the most powerful technique in diagnostic clinical medicine and biomedical researches. With the development of molecular imaging responsive probes allowing multimodal imaging of molecular events are then required.In this work, we present the design and the synthesis of lanthanide complexes with the aim of developing smart contrast agents for the detection of enzyme activity by MRI (T1 / CEST) and Optical Imaging. The designed complexes are built around a macrocyclic lanthanide chelate appended with an amino pyridine which is linked to an enzyme-sensitive trigger (e.g. galactoside) via a self-immolative linker. The latter is supposed to modify temporarily and in an enzyme dependent way the magnetic and photo-physical properties of the complex. The concept was first validated on a model compound without trigger. Although no difference of relaxivity was observed between models of the enzyme-activated and non-activated forms precluding the use in T1-MRI, different paraCEST effects were observed and found dependent on the lanthanide. Moreover, a previously unknown CEST effect was assigned to a carbamate function. Preliminary photo-physical studies showed also a different behavior of the two forms and confirmed the potentiality of these complexes as enzyme responsive bimodal contrast agent. The synthesis of the enzyme-responsive probe has been attempted by three different pathways and was finally achieved in a thirteen-step process which remained to be optimized. A “structure activity” relationship study has been initiated with the synthesis of positional isomers on the pyridine of the model compound

Electrical stimulation of the retina affected by photoreceptor loss (e.g., cases of retinitis pigmentosa) elicits the perception of luminous spots (so-called phosphenes) in the visual field. This phenomenon, attributed to the relatively high survival rates of neurons comprising the retina's inner layer, serves as the cornerstone of efforts to provide a microelectronic retinal prosthesis -- a device analogous to the cochlear implant. This thesis concerns phosphenes -- their elicitation and modulation, and, in turn, image analysis for use in a prosthesis. This thesis begins with a comparative review of visual modelling of electrical epiretinal stimulation and analogous acoustic modelling of electrical cochlear stimulation. The latter models involve coloured noise played to normal listeners so as to investigate speech processing and electrode design for use in cochlear implants. Subsequently, four experiments (three psychophysical and one numerical), and two statistical analyses, are presented. Intrinsic signal optical imaging in cerebral cortex is canvassed appendically. The first experiment describes a visual tracking task administered to 20 normal observers afforded simulated prosthetic vision. Fixation, saccade, and smooth pursuit, and the effect of practice, were assessed. Further, an image analysis scheme is demonstrated that, compared to existing approaches, assisted fixation and pursuit (but not saccade) accuracy (35.8% and 6.8%, respectively), and required less phosphene array scanning. Subsequently, (numerical) information-theoretic reasoning is provided for the scheme's superiority. This reasoning was then employed to further optimise the scheme (resulting in a filter comprising overlapping Gaussian kernels), and may be readily extended to arbitrary arrangements of many phosphenes. A face recognition study, wherein stimuli comprised either size- or intensity-modulated phosphenes, is then presented. The study involved unpracticed observers (n=85), and showed no 'size' --versus--'intensity' effect. Overall, a 400-phosphene (100-phosphene) image afforded subjects 89.0% (64.0%) correct recognition (two-interval forced-choice paradigm) when five seconds' scanning was allowed. Performance fell (64.5%) when the 400-phosphene image was stabilised on the retina and presented briefly. Scanning was similar in 400- and 100-phosphene tasks. The final chapter presents the statistical effects of sampling and rendering jitter on the phosphene image. These results may generalise to low-resolution imaging systems involving loosely packed pixels.

This thesis develops and evaluates three approaches for quantitative molecularly-targeted optical imaging of neoplasia. The first approach focuses on widefield imaging of biomarkers near the tissue surface for early detection applications; this approach is demonstrated in freshly resected oral tissue. Most oral cancers are not detected until the disease has spread, but topical application of targeted imaging agents allows rapid visualization of biomarker expression, giving real-time, objective information. Epidermal growth factor receptor (EGFR) expression was quantified in patient samples using fluorescent epidermal growth factor. Dysplasia (n=4) and cancer (n=13) had an average 2.3-fold and 3.8-fold increase in signal compared to normal tissue. EGFR expression was assessed along with metabolic activity using a fluorescent glucose analog, 2-NBDG, in 9 patient samples. A classification algorithm using quantitative image features resulted in an area under the curve (AUC) of 0.83, though the main advantage of this technique may be to understand spatial heterogeneity of biomarker expression and how this correlates with disease. The next approach focuses on high-resolution optical imaging through a needle to detect metastases in lymphoid tissue for clinical staging applications; this approach is demonstrated in resected lymph nodes from breast cancer patients. These patients often require removal of nodes, but an optical imaging strategy using topical application of imaging agents in vivo may classify nodes as normal or metastatic, thus reducing unnecessary removal of normal nodes and improving metastasis detection. Proflavine, a nuclear dye, was topically applied to 43 nodes. A classification algorithm developed from quantitative image features distinguished normal lymphoid tissue from metastases with an AUC of 0.84. Because optical imaging is depth limited, the final approach combines high-resolution optical imaging with magnetic resonance imaging (MRI) for multimodal evaluation of deep tissue. An imaging agent functional in both optical and MRI was developed by co-loading fluorescent EGFR antibodies and gadolinium-based contrast agents in silicon discs. These discs accumulate in tumors, resulting in localized delivery of imaging agents. The research presented here can be applied to understanding tumor biology and biomarker heterogeneity, with the future clinical goal of improving identification of disease and determination of appropriate therapy for cancer patients.