<p>Worldwide, head and neck squamous cell cancers (HNSCC) account for over 375,000 deaths annually. The majority of these cancers arise in the outermost squamous cells which progress through a series of precancerous changes before developing into invasive HNSCC. It is widely accepted that prognosis is strongly correlated to the stage of diagnosis, with early detection more than doubling the patient’s chance of survival. Currently, however, 60% of HNSCCs are diagnosed when they have already progressed to stage 3 or stage 4 disease. The current diagnostic method of visual examination often fails to recognize early indicators of HNSCC, thereby missing an important prevention window.</p><p> </p><p>Determination of cancer from non-malignant tissues is dependent on pathological examination of lesion biopsies. Thus, all patients with any clinically suspicious lesions undergo surgical biopsies. Furthermore, these surgical biopsies carry risks. In addition to the risk of general anesthesia for patients undergoing panedoscopy, some patients have poor healing and develop ulcerations or infections as a result of surgical biopsy at any anatomical site. Additionally, studies have shown that approximately 50% of suspected biopsies are later pathologically confirmed normal. An enormous amount of labor, facility, and monetary resources are expended on non-malignant biopsies and patients who ultimately have no malignancy. It would be of immense overall benefit to clinicians and patients to have a non-invasive and portable technique that could rapidly identify those patients that would benefit from further surgical biopsy from those that only need follow-up clinical observations.</p><p> </p><p>Once carcinoma is confirmed in a patient, treatment currently involves modalities of surgery, radiation, and chemotherapy. Radiotherapy plays a significant role, particularly in the management of localized HNSCC, because it is a non-invasive and function-preserving modality. However, the effectiveness of radiotherapy is limited by hypoxia. Previous studies showed that tumors reoxygenated during radiotherapy treatment may have a better prognosis. Despite decades of work, there is still no reliable, cost-effective way for measuring tumor hypoxia over multiple time points to estimate the prognosis. </p><p>To address these unmet clinical needs, three aims were proposed. The first aim was to improve early detection by identifying biomarkers of early pre-cancer as well as developing an objective algorithm to detect early disease. Neovasculature is an important biomarker for early cancer diagnosis. Even before the development of a clinically detectable lesion, the tumor vasculature undergoes structural and morphological changes in response to oncogenic signaling pathways [8]. Without receiving a sufficient supply of oxygen and nutrients to proliferate, early tumor growth is limited to only 1-2 mm. High-resolution optical imaging is well suited to characterize the earliest neovascularization changes that accompany neoplasia owing to its sensitivity to hemoglobin absorption and resolution to visualize capillary level architecture. Dark field microscopy is a low-cost and robust method to image the neovasculature. We imaged neovascularization in vivo in a spontaneous hamster oral mucosa carcinogen model using a label-free, reflected-light spectral dark field microscope. Hamsters’ cheek pouches were painted with 7, 12-Dimethylbenz[a]anthracene (DMBA) to induce precancerous to cancerous changes, or mineral oil as control. Spectral dark field images were obtained during carcinogenesis and in control oral mucosa, and quantitative vascular features were computed. Vascular tortuosity increased significantly in oral mucosa diagnosed as hyperplasia, dysplasia and squamous cell carcinoma (SCC) compared to normal. Vascular diameter and area fraction decreased significantly in dysplasia and SCC compared to normal. The areas under the receiver operative characteristic (ROC) curves (AUC) computed using a Support Vector Machine (SVM) were 0.95 and 0.84 for identifying SCC or dysplasia, respectively, vs. normal and hyperplasia oral mucosa combined. To improve AUCs for identifying dysplasia, quantitative vascular features were computed again after the vessels were split into large and small vessels based on diameter. The large vessels preserved the same significant trends, while small vessels demonstrated the opposite trends. Significant increases in diameter and decreases in area fraction were observed in SCC and dysplasia. The AUCs were improved to 0.99 and 0.92 for identifying SCC and dysplasia. These results suggest that dark field vascular imaging is a promising tool for pre-cancer detection.</p><p>Optical imaging can also be applied to quantifying other important characteristics of solid tumors in head and neck cancer (HNC), such as hypoxia, abnormal vascularity and cell proliferation. Diffuse reflectance spectroscopy is a simple and robust method to measure tissue oxygenation, vascularity and cell density. It is particularly suitable for applications in the operation room because of its compact design and portability. In addition, a fiber probe-based system is ideal for obtaining measurements at suspicious lesions in the head and neck area during surgery. Thus, my second aim was to reduce the number of unnecessary HNSCC biopsies by developing a robust tool and rapid analysis method appropriate for clinical settings. We propose the use of morphological optical biomarkers for rapid detection of human HNSCC by leveraging the underlying tissue characteristics in the aerodigestive tracts Prior to biopsy, diffuse reflectance spectra were obtained from malignant and contra-lateral non-malignant tissues of 57 patients undergoing panendoscopy. Oxygen saturation (SO2), total hemoglobin concentration ([THb]), and the reduced scattering coefficient were extracted using an inverse Monte Carlo (MC) method previously developed by former student in our lab. Differences in malignant and non-malignant tissues were examined based on two different groupings: by anatomical site and by morphological tissue type. Measurements were acquired from 252 sites, 51 of which were pathologically classified as SCC. Optical biomarkers exhibited statistical differences between malignant and non-malignant samples. Contrast was enhanced when parsing tissues by morphological classification rather than by anatomical subtype for unpaired comparisons. Corresponding linear discriminant models using multiple optical biomarkers showed improved predictive ability when accounting for morphological classification, particularly in node-positive lesions. The false-positive rate was retrospectively found to decrease by 34.2% in morphologically- vs. anatomically-derived predictive models. In glottic tissue, the surgeon exhibited a false-positive rate of 45.7% while the device showed a lower false-positive rate of only 12.4%. Additionally, comparisons of optical parameters were made to further understand the physiology of tumor staging and potential causes of high surgeon false-positive rates. Optical spectroscopy is a user-friendly, non-invasive tool capable of providing quantitative information to discriminate malignant from non-malignant head and neck tissues. Predictive models demonstrated promising results for diagnostics. Furthermore, the strategy described appears to be well suited to reduce the clinical false-positive rate.</p><p>To further improve the speed for extracting the tissue oxygenation and [THb] to reduce the time when patients were under anesthesia, the third aim was to develop a rapid heuristic ratiometric analysis for estimating tissue [THb] and SO2 from measured tissue diffuse reflectance spectra. The analysis was validated in tissue-mimicking phantoms and applied to clinical measurements in head and neck, cervical and breast tissues. The analysis works in two steps. First, a linear equation that translates the ratio of the diffuse reflectance spectra at 584 nm to 545 nm to estimate the tissue [THb] using a Monte carlo (MC)-based lookup table was developed. This equation is independent of tissue scattering and oxygen saturation. Second, SO2 was estimated using non-linear logistic equations that translate the ratio of the diffuse reflectance spectra at 539 nm to 545 nm into the tissue SO2. Correlations coefficients of 0.89 (0.86), 0.77 (0.71) and 0.69 (0.43) were obtained for the tissue hemoglobin concentration (oxygen saturation) values extracted using the full spectral MC and the ratiometric analysis, for clinical measurements in head and neck, breast and cervical tissues, respectively. The ratiometric analysis was more than 4000 times faster than the inverse MC analysis for estimating tissue [THb] and SO2 in simulated phantom experiments. In addition, the discriminatory power of the two analyses was similar. These results show the potential of such empirical tools to rapidly estimate tissue hemoglobin and oxygenation for real-time applications.</p><p>In addition to its use as a diagnostic marker for various cancers, tissue oxygenation is believed to play a role in the success of cancer therapies, particularly radiotherapy. However, since little effort has been made to develop tools to exploit this relationship, the fourth aim was to estimate patient prognosis by measuring tumor hypoxia over multiple time points so physicians are able to develop more informed and effective clinical treatment plan. To test if oxygenation kinetics correlates with the likelihood for local tumor control following fractionated radiotherapy, we again used diffuse reflectance spectroscopy to noninvasively measure tumor vascular oxygenation and [THb] associated with radiotherapy of 5 daily fractions (7.5, 9 or 13.5 Gy/day) in FaDu xenografts. Spectroscopy measurements were obtained immediately before each daily radiation fraction and during the week after radiotherapy. SO2 and [THb] were computed using an inverse MC model. Oxygenation kinetics during and after radiotherapy, but before a change in tumor volume, was associated with local tumor control. Locally controlled tumors exhibited significantly faster increases in oxygenation after radiotherapy (days 12-15) compared with tumors that recurred locally. (2) Within the group of tumors that recurred, faster increases in oxygenation during radiotherapy (days 3-5) were correlated with earlier recurrence times. An AUC of 0.74 was achieved when classifying the local control tumors from all irradiated tumors using the oxygen kinetics with a logistic regression model. (3) The rate of increase in oxygenation was radiation dose dependent. Radiation doses ≤9.5 Gy/day did not initiate an increase in oxygenation whereas 13.5 Gy/day triggered significant increases in oxygenation during and after radiotherapy. Additional confirmation is required in other tumor models, but these results suggest that monitoring tumor oxygenation kinetics could aid in the prediction of local tumor control after radiotherapy.</p><p>Angiogenesis is a highly regulated process to support tissue growth. Neovasculature is designed by nature to grow toward areas lacking nutrition and oxygen. Cancer cells proliferate too quickly to have their nutritional and oxygen needs completely satisfied, which results in an imbalanced state of angiogenesis leading to tortuous blood vessels, hypoxic tissues and radioresistance. We characterized the tumor-induced vascular features with simple, robust and low-cost dark field microscopy and spectroscopy to enable early cancer diagnosis, improvement of surgical biopsy accuracy and better predict the prognosis of radiotherapy for HNC. Our results demonstrated that these noninvasively measured, label-free vascular features are able to detect pre-cancer, reduce unnecessary surgical biopsies and predict prognosis of radiotherapy.</p> / Dissertation

L'estimation de la profondeur des veines sous-cutanées a été un sujet de recherche important ces dernières années en raison de son importance dans l'optimisation de pose de cathéters, de perfusions et plus généralement de ponctions veineuses. Par le passé, diverses techniques et systèmes de visualisation des veines ont été proposés, cependant le manque d'information sur la profondeur de la veine limite les possibilités pour une automatisation de la ponction veineuse ; le geste clinique restant dans de nombreux cas tributaire des compétences ou de l'expérience des cliniciens. Plusieurs techniques ont été proposées pour estimer la profondeur de la veine en utilisant la réflectance diffuse dont le principe repose sur la mesure de rapport de densité optique (ODR). Le concept de mesure de la profondeur des veines à l'aide de la technique ODR mérite d'être appliqué dans le monde réel en raison de son faible coût, de ses propriétés non invasives et du fait qu'il s'agit d'une technique de mesure sans contact avec la peau. Les travaux initiaux de Nishidate et. Al. [1] ont montré sur fantôme des résultats prometteurs. Cependant, une telle expérience peut ne pas être suffisante pour prouver son application pour la mesure in vivo en raison du manque d'expérience pour les données réelles. Par conséquent, ce travail de thèse a été commencé pour améliorer le modèle proposé par Nishidate et. Al. et l'élargir pour mesurer l'estimation in vivo de la profondeur de la veine sur de vrais patients. Le système proposé intègre de nouveaux composants tels qu'un algorithme de segmentation des veines, une méthode d'estimation plus précise du contenu en mélanine (Cm) et une conception matérielle entièrement nouvelle avec des composants stables. Les résultats obtenus par ODR ont été comparés à des données fournies par une machine Ultrason médicale. Les résultats de l'expérience montrent une corrélation de Pearson forte de 0,843 par rapport aux données échographiques et prouvent que le système développé est fiable pour la mesure in vivo de la profondeur de la veine. En outre, il est proposé d'utiliser un filtre de segmentation de veine optimal (filtre adapté) dans le système d'imagerie pour permettre une segmentation et par la suite une mesure de la profondeur automatique. / The estimation of subcutaneous vein depth has been an important research topic in recent years due to its importance in optimizing the intravenous (IV) access of venipuncture. Various techniques and system of vein visualization were proposed to improve the vein viewing, but the lack of vein depth information limits the system performance in assisting the IV access; thus, the IV access in many cases remains dependent on skill or experience of the clinicians. Several techniques were proposed to estimate the vein depth using diffuse reflectance of which the optical density ratio (ODR) technique is the most complete solution. The concept of measuring the veins depth using ODR based technique is deserved to be applied in the real-world due to its low cost, non-invasive properties and from the fact that it is a non-skin contact measurement technique. Nishidate et. al. [1] suggested an optimum conditions to measure the vein depth and thickness by using ODR which was supported by experiment with customized tissue-like agar gel phantom. However, such experiment may not be sufficient to prove its application for in vivo measurement due to the lack of experiment for real data. Therefore, this thesis work was first started to improve the proposed model by Nishidate et. al. and expand it to measure the in vivo estimation of vein depth on real patients. The proposed system incorporates new components such as an autonomous vein segmentation algorithm, a more accurate estimation method for melanin content (Cm) and a fully new hardware design with reliable parts. Importantly, the experiment estimate the vein depth on real patients as well as a through comparison with Ultrasound data. The experiment results show a strong Pearson correlation of 0.843 as compared to Ultrasound data and this evidence that the developed system is works for the in vivo measurement of vein depth. Besides that, an optimum vein filter (matched filter) is proposed to be used in the imaging system to preserve the most accurate vein detection and allow the system to produce the results with least detection error. The selection of the optimum vein filter has laid an important platform from which to obtain the accurate vein segmentation of a NIR image.

Dentre as diversas aplicações da luz em nosso dia-a-dia, as ligadas à área biomédica merecem destaque e são frequentemente objetos de pesquisa tanto para o desenvolvimento quanto para o aprimoramento de técnicas para o diagnóstico e terapias. Os tecidos biológicos são, em sua maioria, estruturas complexas, não-homogêneas e opticamente muito espalhadoras. Apesar das centenas de estudos existentes acerca da propagação da luz em tecidos biológicos, sua complexidade exige que novos estudos sejam conduzidos a fim de aprimorar o conhecimento já existente, que ainda apresenta muitas lacunas. A presença de heterogeneidades nos tecidos (vasos sanguíneos, hematomas, cistos, tumores e outras alterações macroscópicas) mudam a propagação da luz e dificultam a previsibilidade do seu comportamento por modelos matemáticos. O presente trabalho teve por objetivo estabelecer um método empírico que utiliza imagens de refletância difusa obtidas através de uma instrumentação simples, baseada em uma fonte de luz contínua no visível (laser de diodo em 660 nm) e uma câmera CMOS monocromática, para verificar a possibilidade de localização de inclusões absorvedoras embebidas em phantoms altamente espalhadores. Foi avaliada a capacidade de detectar inclusões de dois diferentes tipos de materiais em diversas geometrias e tamanhos, posicionadas em diferentes profundidades. O ângulo de incidência do feixe laser também foi variado, bem como a distância entre a fonte e o objeto, a fim de avaliar quais as melhores condições experimentais. Os resultados obtidos mostraram que os objetos puderam ser detectados, e suas formas puderam ser satisfatoriamente recuperadas através de um algoritmo desenvolvido para o processamento das imagens. Em algumas situações, mesmo para a maior profundidade utilizada, que foi de 20 mm, a inclusão pôde ser detectada nas imagens de refletância difusa processadas. Apesar da capacidade de detecção das formas geométricas representar um avanço com relação às possibilidades de identificação de estruturas em meios túrbidos, a determinação da profundidade ainda é um desafio a ser superado. / Among the various applications of light in our daily life, those connected with biomedicine should be highlighted and are frequently subject of researches aiming for the development and for the enhancement of techniques for diagnosis and therapy. Biological tissues are mostly complex, non-homogeneous and optically highly scattering structures. Despite the hundreds of existent studies on the propagation of light in biological tissues, its complexity requires new studies to be conducted in order to improve the existing knowledge, which still has many gaps. The presence of heterogeneities in tissue (blood vessels, bruises, cysts, tumors and other macroscopic alteration) changes the light propagation and impedes the predictability of its behavior by mathematical models. This work aimed to establish an empirical method using diffuse reflectance images acquired with simple instrumentation, based on a source of continuous light in the visible (diode laser at 660 nm) and a monochromatic CMOS camera, to check the possibility of the location of absorbing inclusions embedded in highly scattering phantoms. The ability to detect inclusions of two different kinds of materials in different sizes and geometries, positioned at different depths were evaluated. The laser beam angle of incidence was also varied, as well as the distance between the source and the object, in order to evaluate the best experimental conditions. The results showed that the objects could be detected, and their shapes might be satisfactorily recovered by an algorithm developed for image processing. In some situations, even at the greatest depth used, which was 20 mm, the inclusion could be detected in diffuse reflectance processed images. Although the detection capability of geometric shapes represents an improvement over the structures of identification possibilities in turbid media, the determination of depth is still a challenge to be overcome.

O principal parâmetro de avaliação da qualidade de carvões para uso na geração de energia termoelétrica é o percentual de cinzas. Em uma planta de beneficiamento, saber o percentual de cinzas da alimentação, concentrado e rejeito é de suma importância para os ajustes nos equipamentos de concentração. No caso do carvão da camada I1 do depósito mineral do Leão a janela de tempo entre o ponto mais adequado para amostragem e o fim da campanha de beneficiamento não propicia que se conheça o teor de cinzas da alimentação do processo de beneficiamento. Em virtude disso, este trabalho tem como objetivo de fornecer uma estimativa confiável do teor de cinzas da alimentação para propiciar a aplicação na planta de beneficiamento. Para esse intento utilizou-se a teoria de Kubelka-Munk que por meio de sua expressão matemática transforma a reflectância em uma grandeza aditiva que propicia a estimativa das porcentagens de matéria orgânica e mineral contida no carvão devido a diferença de reflectância entre esses materiais. Como resultado final foram gerados diversos modelos com intuito de verificar qual deles se adaptava melhor aos dados. Por fim, foi possível concluir que há possibilidade de estimar o teor de cinzas de carvão por meio da reflectância e com confiabilidade de 96,78% que toda a leitura agrega um erro máximo de ± 5% para o melhor modelo. / The main parameter in quality evaluation of coals employed in power generation is the ash content. In a beneficiation plant, knowing the ash content of the feed, concentrate an tailings is mandatory to set up the concentration equipment. In the case of the I1 coal seam of Leão’s deposit the time window between sampling routine and beneficiation routine does not allow to know the feed ash content before the beneficiation process. By this reason, this work aims to provide a reliable estimative of the feed ash content to propitiate its application in the beneficiation plant. For this purpose it was used Kubelka-Munk’s theory that, by using a mathematic expression, transforms reflectance into a additive quantity which allows to estimate mineral and organic matter present in coal by the difference of reflectance between these materials. As result, it was generated several models in order to analyze which one is better suited to the data. Finally, as a conclusion, it is possible to infer the ash content of coal by reflectance with confidence of 96,78% that all measure brings up a maximum error of ± 5% for the best model.

<p>Breast conserving surgery (BCS) is a common treatment option for breast cancer patients. The goal of BCS is to remove the entire tumor from the breast while preserving as much normal tissue as possible for a better cosmetic outcome after surgery. Specifically, the excised specimen must have at least 2 mm of normal tissue surrounding the diseased mass. Unfortunately, a staggering 20-70% of patients undergoing BCS require repeated surgeries due to the incomplete removal of the tumor diagnosed post-operatively. Due to these high re-excision rates as well as limited post-operative histopathological sampling of the tumor specimen, there is an unmet clinical need for margin assessment. Quantitative diffuse reflectance spectral imaging has previously been explored as a promising, method for providing real-time visual maps of tissue composition to help surgeons determine breast tumor margins to ensure the complete removal of the disease during breast conserving surgery. We have leveraged the underlying sources of contrast in breast tissue, specifically total hemoglobin content, beta-carotene content, and tissue scattering, and developed various fiber optics based spectral imaging systems for this clinical application. Combined with a fast inverse Monte Carlo model of reflectance, previous studies have shown that this technology may be able to decrease re-excision rates for BCS. However, these systems, which all consist of a broadband source, fiber optics probes, an imaging spectrograph and a CCD, have severe limitations in system footprint, tumor area coverage, and speed for acquisition and analysis. The fiber based spectral imaging systems are not scalable to smaller designs that cover a large surveillance area at a very fast speed, which ultimately makes them impractical for use in the clinical environment. The objective of this dissertation was to design, develop, test, and show clinical feasibility of a novel wide field spectral imaging system that utilizes the same scientific principles of previously developed fiber optics based imaging systems, but improves upon the technical issues, such as size, complexity, and speed,to meet the demands of the intra-operative setting. </p><p>First, our simple re-design of the system completely eliminated the need for an imaging spectrograph and CCD by replacing them with an array of custom annular photodiodes. The geometry of the photodiodes were designed with the goal of minimizing optical crosstalk, maximizing SNR, and achieving the appropriate tissue sensing depth of up to 2 mm for tumor margin assessment. Without the imaging spectrograph and CCD, the system requires discrete wavelengths of light to launch into the tissue sample. A wavelength selection method that combines an inverse Monte Carlo model and a genetic algorithm was developed in order to optimize the wavelength choices specifically for the underlying breast tissue optical contrast. The final system design consisted of a broadband source with an 8-slot filter wheel containing the optimized set of wavelength choices, an optical light guide and quartz light delivery tube to send the 8 wavelengths of light in free space through the back apertures of each annular photodiode in the imaging array, an 8-channel integrating transimpedance amplifier circuit with a switch box and data acquisition card to collect the reflectance signal, and a laptop computer that controls all the components and analyzes the data.</p><p>This newly designed wide field spectral imaging system was tested in tissue-mimicking liquid phantoms and achieved comparable performance to previous clinically-validated fiber optics based systems in its ability to extract optical properties with high accuracy. The system was also tested in various biological samples, including a murine tumor model, porcine tissue, and human breast tissue, for the direct comparison with its fiber optics based counterparts. The photodiode based imaging system achieved comparable or better SNR, comparable extractions of optical properties extractions for all tissue types, and feasible improvements in speed and coverage for future iterations. We show proof of concept in performing fast, wide field spectral imaging with a simple, inexpensive design. With a reduction in size, cost, number of wavelengths used, and overall complexity, the system described by this dissertation allows for a more seamless scaling to higher pixel number and density in future iterations of the technology, which will help make this a clinically translatable tool for breast tumor margin assessment.</p> / Dissertation

<p>Among women worldwide, cervical cancer is the third most common cancer with an incidence rate of 15.3 per 100,000 and a mortality rate of 7.8 per 100,000 women. This is largely attributed to the lack of infrastructure and resources in the developing countries to support the organized screening and diagnostic programs that are available to women in developed nations. Hence, there is a critical global need for a screening and diagnostic paradigm that is effective in low-resource settings. Various strategies are described to design an optical spectroscopic sensor capable of collecting reliable diffuse reflectance data to extract quantitative optical contrasts for cervical cancer screening and diagnosis. </p><p> A scalable Monte Carlo based optical toolbox can be used to extract absorption and scattering contrasts from diffuse reflectance acquired in the cervix in vivo. [Total Hb] was shown to increase significantly in high-grade cervical intraepithelial neoplasia (CIN 2+), clinically the most important tissue grade to identify, compared to normal and low-grade intraepithelial neoplasia (CIN 1). Scattering was not significantly decreased in CIN 2+ versus normal and CIN 1, but was significantly decreased in CIN relative to normal cervical tissues. </p><p> Immunohistochemistry via anti-CD34, which stains the endothelial cells that line blood vessels, was used to validate the observed absorption contrast. The concomitant increase in microvessel density and [total Hb] suggests that both are reactive to angiogenic forces from up-regulated expression of VEGF in CIN 2+. Masson's trichrome stain was used to assess collagen density changes associated with dysplastic transformation of the cervix, hypothesized as the dominant source of decreased scattering observed. Due to mismatch in optical and histological sampling, as well as the small sample size, collagen density and scattering did not change in a similar fashion with tissue grade. Dysplasia may also induce changes in cross-linking of collagen without altering the amount of collagen present. Further work would be required to elucidate the exact sources of scattering contrast observed. </p><p> Common confounding variables that limit the accuracy and clinical acceptability of optical spectroscopic systems are calibration requirements and variable probe-tissue contact pressures. Our results suggest that using a real-time self-calibration channel, as opposed to conventional post-experiment diffuse reflectance standard calibration measurements, significantly improved data integrity for the extraction of scattering contrast. Extracted [total Hb] and scattering were also significantly associated with applied contact probe pressure in colposcopically normal sites. Hence, future contact probe spectroscopy or imaging systems should incorporate a self-calibration channel and ensure spectral acquisition at a consistent contact pressure to collect reliable data with enhanced absorption and scattering contrasts. </p><p> Another method to enhance optical contrast is to selectively interrogate different depths in the dysplastic cervix. For instance, scattering has been shown to increase in the epithelium (increase in nuclear-to-cytoplasmic ratio) while decrease in the stroma (re-organization of the extra-cellular matrix and changes in of collagen fiber cross-links). A fiber-optic probe with 45° illumination and collection fibers with a separation distance of 330 &#956;m was designed and constructed to selectively interrogate the cervical epithelium. Mean extraction errors from liquid phantoms with optical properties mimicking the cervical epithelium for &#956;a and &#956;s' were 11.3 % and 12.7 %, respectively. Diffuse reflectance spectra from 9 sites in four loop electrosurgical excision procedure (LEEP) patients were analyzed. Preliminary data demonstrate the utility of the oblique fiber geometry in extracting scattering contrast in the cervical epithelium. Further work is needed to study the systematic error in optical property extraction and to incorporate simultaneous extraction of epithelial and stromal contrasts using both flat and oblique illumination and collection fibers. </p><p> Various strategies, namely self-calibration, consistent contact pressure, and the incorporation of depth-selective sensing, have been proposed to improve the data integrity of an optical spectroscopic system for maximal contrast. In addition to addressing field operation requirements (such as power and operator training requirement), these improvements should enable the collection of reliable spectral data to aid in the adoption of optical smart sensors in the screening and diagnosis of cervical precancer, especially in a global health setting.</p> / Dissertation

Ultrathin fiberscopes typically have an imaging channel and an illumination channeland are available in diameters ranging from 0.5 mm to 2.5 mm. The minimum diam-eter can be reduced by combining the illumination and imaging paths into a singlefiberoptic channel. Constructing a single channel fiberscope requires a technique ofilluminating the tissue, while minimizing the Fresnel reflections and scatter withinthe common illumination and detection channel.A single channel fiberscope should image diffusely reflected light from tissue illu-minated with light filtered for the visible wavelength range (450 - 650 nm). Simplycombining the illumination and collection paths via a beamsplitter results in a lowobject to background signal ratio. The low contrast image is due to a low collectionefficiency of light from the ob ject as well as a high background signal from the Fresnelreflection at the proximal surface of the fiber bundle, where the illumination enters thefiber bundle. The focus of the dissertation is the investigation of methods to reducethe background signal from the proximal surface of the fiber bundle. Three systemswere tested. The first system uses a coherent fiber bundle with an AR-coating on theproximal face. The second system incorporates crossed polarizers into the light path.In addition, a technique was developed, whereby a portion of the image numericalaperture is devoted to illumination and a portion to image signal detection. Thistechnique is called numerical aperture sharing (NA sharing).This dissertation presents the design, construction, testing, and comparison ofthe three single channel fiberscopes. In addition, preliminary results of a study aimedat the usefulness of broadband diffuse reflectance imaging for the identification andtracking of disease progression in mouse esophagus are presented.

Resumo: O presente trabalho propõe o desenvolvimento de um método limpo para determinação de ureia em amostras comerciais de fertilizantes utilizando a Espectroscopia de Reflectância Difusa combinada com spot test. O método desenvolvido é baseado na reação entre a ureia e o reagente cromogênico p-dimetilaminocinamaldeído (p-DAC) em meio de ácido clorídrico diluído, que resulta em um produto de coloração rósea cujo valor máximo de reflectância ocorre no comprimento de onda de 535 nm. Os parâmetros experimentais foram otimizados através dos planejamentos fatorial e composto central para a obtenção da superfície de resposta, os quais indicaram uma maior sensibilidade para o método quando se utiliza o reagente p-DAC na concentração de 0,196% (m/v) em meio de ácido clorídrico de concentração 0,0549 mol L-1, empregando etanol como solvente para as soluções. O suporte sólido para a reação é um papel de filtro qualitativo onde são colocados 20,0μL de solução de ureia seguidos de secagem com ar frio de um secador de cabelos e 20,0μL de solução de reagente p-DAC 0,196% (m/v) em meio de HCl 0,0549 mol L-1 e secagem ao ar livre, submetendo-se então o spot à leitura reflectométrica em máx. = 535 nm. Uma relação linear (R = 0,996) foi estabelecida na faixa de concentração de ureia compreendida entre 50,0 - 300 mg L-1. Os limites de detecção e de quantificação foram determinados em 5,13 mg L-1 e 17,10 mg L-1, respectivamente. O método limpo desenvolvido foi aplicado na determinação de ureia em amostras comerciais de fertilizantes onde demonstrou apresentar ótima precisão e exatidão, evidenciadas pela boa recuperação (94,2 - 107,4%), além de oferecer vantagens como simplicidade de execução e baixo consumo de reagentes, geração de mínima quantidade de resíduos, rapidez e segurança... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This work proposes the development of a clean method for determination of urea in commercial samples of fertilizers using the Diffuse Reflectance Spectroscopy combined with spot test. The method is based on the reaction between urea and the chromogenic reagent p-dimethylaminocinnamaldehyde (p-DAC) in diluted hydrochloric acid medium, which results in a pink colored product whose maximum value of reflectance occurs at a wavelength of 535 nm. The experimental parameters were optimized through a factorial and a central composite experimental design to obtain the response surface, which indicated a higher sensitivity for the method when using the reagent p-DAC in the concentration of 0.196% (w/v) in a hydrochloric acid 0.0549 mol L-1 medium, using ethanol as a solvent for the solutions. The solid support for the reaction is a qualitative filter paper where they are placed 20.0μL of urea solution followed by drying with cold air from a hair dryer and 20.0μL of reagent solution p-DAC 0.196% (w/v) in the HCl 0.0549 mol L-1 medium and drying in the air, then submitting the spot to the reflectometric reading at max = 535 nm. A linear relationship (R = 0.996) was established in the range of urea concentration between 50.0 to 300 mg L-1. The limits of detection and quantification were determined at 5.13 mg L-1 and 17.10 mg L-1, respectively. The method was applied to the clean determination of urea in commercial samples of fertilizers, which has shown to present great precision and accuracy as evidenced by the good recovery (94.2 to 107.4%) and also offers advantages such as simplicity of implementation and low consumption of reagents, generation of minimum quantity of wastes, rapidity and security, producing reliable results. As shown, the method is designed as environmentally friendly for determination of urea, as it is consistent with the Principles advocated by the Green Chemistry. / Orientador: Helena Redigolo Pezza / Coorientador: Leonardo Pezza / Banca: Maria Del Pilar Taboada Sotomayor / Banca: Adriana Vitorino Rossi / Mestre

O principal parâmetro de avaliação da qualidade de carvões para uso na geração de energia termoelétrica é o percentual de cinzas. Em uma planta de beneficiamento, saber o percentual de cinzas da alimentação, concentrado e rejeito é de suma importância para os ajustes nos equipamentos de concentração. No caso do carvão da camada I1 do depósito mineral do Leão a janela de tempo entre o ponto mais adequado para amostragem e o fim da campanha de beneficiamento não propicia que se conheça o teor de cinzas da alimentação do processo de beneficiamento. Em virtude disso, este trabalho tem como objetivo de fornecer uma estimativa confiável do teor de cinzas da alimentação para propiciar a aplicação na planta de beneficiamento. Para esse intento utilizou-se a teoria de Kubelka-Munk que por meio de sua expressão matemática transforma a reflectância em uma grandeza aditiva que propicia a estimativa das porcentagens de matéria orgânica e mineral contida no carvão devido a diferença de reflectância entre esses materiais. Como resultado final foram gerados diversos modelos com intuito de verificar qual deles se adaptava melhor aos dados. Por fim, foi possível concluir que há possibilidade de estimar o teor de cinzas de carvão por meio da reflectância e com confiabilidade de 96,78% que toda a leitura agrega um erro máximo de ± 5% para o melhor modelo. / The main parameter in quality evaluation of coals employed in power generation is the ash content. In a beneficiation plant, knowing the ash content of the feed, concentrate an tailings is mandatory to set up the concentration equipment. In the case of the I1 coal seam of Leão’s deposit the time window between sampling routine and beneficiation routine does not allow to know the feed ash content before the beneficiation process. By this reason, this work aims to provide a reliable estimative of the feed ash content to propitiate its application in the beneficiation plant. For this purpose it was used Kubelka-Munk’s theory that, by using a mathematic expression, transforms reflectance into a additive quantity which allows to estimate mineral and organic matter present in coal by the difference of reflectance between these materials. As result, it was generated several models in order to analyze which one is better suited to the data. Finally, as a conclusion, it is possible to infer the ash content of coal by reflectance with confidence of 96,78% that all measure brings up a maximum error of ± 5% for the best model.

Submitted by LAÉRCIO SANTOS SILVA null (laerciosantos18@gmail.com) on 2016-03-21T21:18:03Z No. of bitstreams: 1 Dissertação_Laercio_Santos_Silva_espelhada_2016.pdf: 1714650 bytes, checksum: 47f05faa577cbb7bf53e0c788cea097a (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-03-22T19:10:43Z (GMT) No. of bitstreams: 1 silva_ls_me_jabo.pdf: 1714650 bytes, checksum: 47f05faa577cbb7bf53e0c788cea097a (MD5) / Made available in DSpace on 2016-03-22T19:10:43Z (GMT). No. of bitstreams: 1 silva_ls_me_jabo.pdf: 1714650 bytes, checksum: 47f05faa577cbb7bf53e0c788cea097a (MD5) Previous issue date: 2016-02-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A diversidade geológica do Planalto condicionou a formação de diversos solos, predominando na fração argila caulinita, óxidos e hidróxidos de ferro e gibbsita. Entre estes, os óxidos e hidróxidos de ferro e a gibbsita são considerados pedoindicadores ambientais na região tropical. Desta forma, os objetivos desta pesquisa foram: i) caracterizar os óxidos de ferro pedogênicos formados a partir de compartimentos geológicos com baixos e altos teores de ferro total do Planalto Ocidental Paulista e associá-los com o grau de dissecação da paisagem; ii) caracterizar os óxidos de ferro dos solos do Planalto Ocidental Paulista por espectroscopia de reflectância difusa. Baseado nas informações geológicas, geomorfológica e na amplitude de variação dos teores de ferro pedogênicos (Fed e Feo). Foram coletadas doze amostras de solo na profundidade de 0,0 – 0,2 m para caracterização mineralógica e espectral. A magnitude dos teores totais obtida pela digestão com ácido sulfúrico evidenciou a diferença do material de origem, geologia, intensidade de pedogênese e geomorfologia. Nos solos da Formação Vale do Rio do Peixe predominaram a mineralogia caulinítica, enquanto na Formação Serra Geral, hematítica. Os atributos granulométricos, químicos e mineralógicos foram influenciados pelo grau de dissecação dos solos do Planalto Ocidental Paulista. O comportamento espectral das regiões em diferentes estágios de dissecação, foram diferentes na faixa espectral dos óxidos (530 a 570 nm) e caulinita (1880 a 2380 nm). A caulinita foi melhor pedoindicador para caracterizar a paisagem em diferentes estágios de dissecação nos solos de arenito, enquanto que a hematita e goethita foram melhores pedoindicadores para caracterizar a paisagem em diferentes estágios de dissecação nos solos basalto. Esse resultado pode auxiliar na compreensão da evolução da paisagem e do solo em outras bacias sedimentares. / The geological diversity of Plateau conditioned the formation of various soils, predominantly kaolinite in the clay fraction, iron oxides and hydroxides and gibbsite. Among these, the oxides and hydroxides of iron and gibbsite are considered environmental pedoindicadores in the tropical region. Thus, aimed to The geological diversity of Plateau conditioned the formation of various soils, predominantly kaolinite in the clay fraction, iron oxides and hydroxides and gibbsite. Among these, the oxides and hydroxides of iron and gibbsite are considered environmental pedoindicadores in the tropical region. Thus, aimed to characterize the oxides and hydroxides of iron (hematite and goethite), kaolinite and gibbsite Western Paulista Plateau and associate them with the landscape of the state of play in different environments with total iron content. Based on geological, geomorphological and amplitude variation of pedogenic iron levels (Fed and Feo). Twelve soil samples were collected at a depth 0.0 to 0.2 m for mineralogical and spectral characterization. The magnitude of the total contents obtained by digestion with sulfuric acid showed the difference in the source material, geology, geomorphology and pedogenesis intensity. In soils Training Fish River Valley predominated kaolinitic mineralogy, while the Serra Geral Formation, hematítica. The granulometric, chemical and mineralogical attributes were influenced by the degree of dissection of the Western Paulista Plateau soil. The spectral behavior of the regions at different stages of dissection were different in the spectral range of oxides (530 to 570 nm) and kaolinite (1880 to 2380 nm). The kaolinite was better environmental indicators to characterize the landscape in different stages of dissection in sandy soils, whereas hematite and goethite were better environmental indicators to characterize the landscape at different stages of dissection in basalt soils. This result may help to understand the evolution of the landscape and soil in other sedimentary basins. the oxides and hydroxides of iron (hematite and goethite), kaolinite and gibbsite Western Paulista Plateau and associate them with the landscape of the state of play in different environments with total iron content. Based on geological, geomorphological and amplitude variation of pedogenic iron levels (Fed and Feo). Twelve soil samples were collected at a depth 0.0 to 0.2 m for mineralogical and spectral characterization. The magnitude of the total contents obtained by digestion with sulfuric acid showed the difference in the source material, geology, geomorphology and pedogenesis intensity. In soils Training Fish River Valley predominated kaolinitic mineralogy, while the Serra Geral Formation, hematítica. The granulometric, chemical and mineralogical attributes were influenced by the degree of dissection of the Western Paulista Plateau soil. The spectral behavior of the regions at different stages of dissection were different in the spectral range of oxides (530 to 570 nm) and kaolinite (1880 to 2380 nm). The kaolinite was better environmental indicators to characterize the landscape in different stages of dissection in sandy soils, whereas hematite and goethite were better environmental indicators to characterize the landscape at different stages of dissection in basalt soils. This result may help to understand the evolution of the landscape and soil in other sedimentary basins.