Support vector machine for optical diagnosis of cancer

We report the application of a support vector machine (SVM) for the development of diagnostic algorithms for optical diagnosis of cancer. Both linear and nonlinear SVMs have been investigated for this purpose. We develop a methodology that makes use of SVM for both feature extraction and classification jointly by integrating the newly developed recursive feature elimination (RFE) in the framework of SVM. This leads to significantly improved classification results compared to those obtained when an independent feature extractor such as principal component analysis (PCA) is used. The integrated SVM-RFE approach is also found to outperform the classification results yielded by traditional Fisher's linear discriminant (FLD)-based algorithms. All the algorithms are developed using spectral data acquired in a clinical in vivo laser-induced fluorescence (LIF) spectroscopic study conducted on patients being screened for cancer of the oral cavity and normal volunteers. The best sensitivity and specificity values provided by the nonlinear SVM-RFE algorithm over the data sets investigated are 95 and 96% toward cancer for the training set data based on leave-one-out cross validation and 93 and 97% toward cancer for the independent validation set data. When tested on the spectral data of the uninvolved oral cavity sites from the patients it yielded a specificity of 85%

Automated Biochemical, Morphological, and Organizational Assessment of Precancerous Changes from Endogenous Two-Photon Fluorescence Images

Multi-photon fluorescence microscopy techniques allow for non-invasive interrogation of live samples in their native environment. These methods are particularly appealing for identifying pre-cancers because they are sensitive to the early changes that occur on the microscopic scale and can provide additional information not available using conventional screening techniques.In this study, we developed novel automated approaches, which can be employed for the real-time analysis of two-photon fluorescence images, to non-invasively discriminate between normal and pre-cancerous/HPV-immortalized engineered tissues by concurrently assessing metabolic activity, morphology, organization, and keratin localization. Specifically, we found that the metabolic activity was significantly enhanced and more uniform throughout the depths of the HPV-immortalized epithelia, based on our extraction of the NADH and FAD fluorescence contributions. Furthermore, we were able to separate the keratin contribution from metabolic enzymes to improve the redox estimates and to use the keratin localization as a means to discriminate between tissue types. To assess morphology and organization, Fourier-based, power spectral density (PSD) approaches were employed. The nuclear size distribution throughout the epithelial depths was quantified by evaluating the variance of the corresponding spatial frequencies, which was found to be greater in the normal tissue compared to the HPV-immortalized tissues. The PSD was also used to calculate the Hurst parameter to identify the level of organization in the tissues, assuming a fractal model for the fluorescence intensity fluctuations within a field. We found the range of organization was greater in the normal tissue and closely related to the level of differentiation.A wealth of complementary morphological, biochemical and organizational tissue parameters can be extracted from high resolution images that are acquired based entirely on endogenous sources of contrast. They are promising diagnostic parameters for the non-invasive identification of early cancerous changes and could improve significantly diagnosis and treatment for numerous patients

Optical microsystem for spectroscopy signals extraction applied to gastrointestinal dysplasia detection

Tese de Doutoramento em Engenharia BiomédicaThe early detection of gastrointestinal cancer, in the dysplastic stage, is essential to increase the patient survival rate. Spectroscopic techniques, particularly diffuse reflectance and fluorescence, can improve the gastrointestinal dysplasia detection, since these techniques can be used to extract biochemical and morphologic information related with the status of a gastrointestinal tissue. Several research groups have developed prototypes for the extraction of diffuse reflectance and fluorescence signals applied to gastrointestinal cancer detection. Despite their advantages associated with the gastrointestinal cancer identification, they have several disadvantages related with the use of complex, high-cost and sophisticate components such as xenon lamps, lasers, monochromators, optical fibers and high quantum efficiency detectors, which may hamper their wide use as well as their huge clinical value. Therefore, it is of utmost importance to develop a low-cost, miniaturized and minimal invasive microsystem for spectroscopic signals extraction. As a result, in this work it is proposed the implementation of a microsystem, which comprises in a single chip, an optical filter system for selection and extraction of the diffuse reflectance and fluorescence signals in relevant spectral bands, a silicon photodiodes matrix (4×4) and its readout electronics, and miniaturized light emitting diodes. The main applications of this microsystem are: its use as a portable device in a surgery room for inspection of total removing of the cancerous or dysplastic tissue; and its integration with the standard endoscopes and colonoscopes using it as an auxiliary, to the physician, in early gastrointestinal cancer detection. Along this thesis, important steps towards that microsystem implementation were achieved. In a first step experimental measurements were performed, with phantoms representative of the main absorbing, scattering and fluorescence properties of gastrointestinal tissues (containing hemoglobin, polystyrene microspheres to represent collagen fibers, and the fluorophores NADH and Carbostyril 124, the latter representing collagen), in order to study the diffuse reflectance and fluorescence typical spectra and their temperature dependence. Moreover, the viability of using only 16 spectral bands (between 350 and 750 nm) for signals extraction was discussed, proving the feasibility of an optical filter system implementation in the final microsystem. Therefore, it were designed, fabricated and characterized 16 MgO/TiO2 and SiO2/TiO2 based thin-film optical filters. Their characterization performed through optical transmittance, selectivity, profilometry and scanning electron microscopy, allowed understanding the deviations between the simulated characteristics and the ones experimentally obtained. Moreover, the optical filters results showed transmittances ranging from 50% to 90% approximately, and a full width half maximum (FWHM) averaging from 11 nm to 20 nm, which fits the required application. The fabricated optical filters had some deviations considering their simulated characteristics, which can be explained by the complexity of the optical filters design, for example, the materials refractive index dependence with wavelength and thin-film thickness. The diffuse reflectance and fluorescence signals that pass through the optical filters can be measured with an on-chip silicon photodetectors matrix (4×4), based on n+/p-epilayer junction photodiodes with an active area of 100 × 100 µm2, and a light-to-frequency converter, per each photodiode, that enables producing a digital signal with a frequency proportional to the photodiode current. As a result, the design and implementation of a CMOS microsystem comprising these components were performed. The photodiodes characterization showed a responsivity of 200 mA/W at 550 nm, approximately, and the light-to-frequency converter connected to the photodiode showed a linear response (R2>0.99) with a sensitivity of 25 Hz/nA at 550 nm, approximately. The behavior of the current-to-frequency converter, with an external current source directly injected in its input, was also studied allowing to confirm its linearity in the range of currents produced in this application, its power consumption of 1 mW, and its maximum input current, approximately 300 µA. This CMOS approach avoids the need of an expensive readout optical microsystem, since it is possible to integrate the photodiodes and the readout electronics in a small silicon area (275 × 100 µm2 per photodiode and its respective converter). The performance of the implemented microsystem and the fabricated optical filters was evaluated, using phantoms (also containing hemoglobin, polystyrene microspheres, NADH and Carbostyril 124). The obtained results have shown the viability of the microsystem (including the optical filter system) to extract diffuse reflectance and fluorescence signals. Some issues were noted on the sensitivity of the implemented optical setups for the on-chip measurements. However, some solutions were proposed for the remaining problems, specifically the future use of miniaturized light emitting diodes and the direct deposition of the optical filters on the top of the photodetection system. Finally, the direct integration of optical filters on top of the photodiodes was discussed and a new approach was tested.The author, Sara Filomena Ribeiro Pimenta, was supported by the Portuguese Foundation for Science and Technology (in portuguese FCT – Fundação para a Ciência e a Tecnologia) with the PhD grant SFRH/BD/87605/2012. This work is also funded by FEDER funds through the “Eixo I do Programa Operacional Fatores de Competitividade” (POFC) QREN, project reference COMPETE: FCOMP-01-0124-FEDER- 020241 and by FCT, project reference PTDC/EBB-EBI/120334/2010. Finally, the author thanks to the PEst-C/FIS/UI0607/2013, UID/FIS/04650/2013, UID/EEA/04436/2013 and POCI-01- 0145-FEDER-006941 for the use of equipment.A deteção precoce do cancro gastrointestinal, na fase de displasia, é essencial para o aumento da taxa de sobrevivência do paciente. As técnicas de espetroscopia, particularmente a refletância difusa e a fluorescência, permitem melhorar a deteção de displasia gastrointestinal, ao poderem ser utilizadas para a extração de informação bioquímica e morfológica associada ao estado do tecido gastrointestinal. Diversos grupos de investigação têm desenvolvido protótipos para a extração de sinais de refletância difusa e de fluorescência, para aplicação na deteção do cancro gastrointestinal. Apesar das vantagens associadas com a identificação do cancro gastrointestinal, esses sistemas apresentam várias desvantagens relacionadas com a utilização de componentes complexos, de elevado custo e sofisticados, como por exemplo, lâmpadas de xénon, lasers, monocromadores, fibras óticas e detetores de elevada eficiência, que podem dificultar a sua ampla utilização, bem como o seu elevado valor clínico. Portanto, é de extrema importância o desenvolvimento de um microssistema de baixo custo, miniaturizado e minimamente invasivo para a extração de sinais de espetroscopia. Assim, neste trabalho é proposta a implementação de um microssistema, num único chip, compreendendo: um sistema de filtros óticos para a seleção dos sinais de refletância difusa e de fluorescência em bandas espetrais relevantes; uma matriz de fotodíodos de silício (4×4) e a respetiva eletrónica de leitura; e díodos emissores de luz miniaturizados. As principais aplicações deste microssistema são: a sua utilização como sistema portátil numa sala de cirurgia para inspeção da remoção total do tecido maligno ou displásico; ou a sua integração com os sistemas de endoscopia e colonoscopia, servindo como auxiliar de diagnóstico, na deteção precoce de cancro gastrointestinal. Com a realização desta tese foram dados passos importantes para a implementação desse microssistema. Numa primeira fase, foram realizados testes experimentais, com um grupo de fantomas representativos das propriedades de absorção, difusão e de fluorescência dos tecidos gastrointestinais (contendo hemoglobina, microesferas de polistireno representando as fibras de colagénio, e os fluoróforos NADH e Carbostyril 124, este último para representar o colagénio), de forma a obter os espetros típicos de refletância difusa e de fluorescência e a influência da temperatura do fantoma nos mesmos. Para além disso, a viabilidade de usar apenas 16 bandas espetrais (entre 350 e 750 nm) para a extração dos sinais espetroscópicos foi discutida, provando a exequibilidade da implementação de um sistema de filtros óticos no microssistema final. Assim, foram desenhados, fabricados e caracterizados 16 filtros óticos baseados em filmes finos de MgO/TiO2 e SiO2/TiO2. A sua caracterização do ponto de vista da transmitância ótica, seletividade, profilometria e microscopia eletrónica de varrimento, permitiu perceber os desvios verificados entre as características simuladas e as obtidas experimentalmente. Para além disso, os resultados da caracterização dos filtros óticos mostraram transmitâncias óticas que variam entre 50% e 90%, aproximadamente, e uma largura a meia-altura (FWHM) média entre 11 nm e 20 nm, o que é adequado para a aplicação pretendida. Os filtros óticos fabricados possuem alguns desvios das suas características simuladas, o que pode ser explicado pela complexidade no projeto de filtros óticos, por exemplo, a dependência dos índices de refração com o comprimento de onda e espessura do filme fino. Os sinais de refletância difusa e fluorescência que atravessam os filtros óticos podem ser medidos através de uma matriz de fotodetetores de silício (4×4), baseada em fotodíodos do tipo n+/p-epilayer com uma área ativa de 100 × 100 µm2, e um conversor luz-frequência, um por cada fotodíodo, que permite produzir um sinal digital com uma frequência proporcional à corrente gerada pelo fotodíodo. Assim, o projeto e a implementação de um microssistema CMOS incluindo esses componentes foram executados. A caracterização dos fotodíodos da matriz resultou num valor de responsividade de 200 mA/W a 550 nm, aproximadamente, e a do conversor luz-frequência, quando ligado a um fotodíodo, resultou numa resposta linear (R2>0.99) com uma sensibilidade de 25 Hz/nA a 550 nm, aproximadamente. O comportamento do conversor corrente-frequência, com uma fonte de corrente externa diretamente injetada na sua entrada, foi também estudado, permitindo confirmar a sua linearidade na gama de correntes envolvidas nesta aplicação, a sua potência de consumo de 1 mW, e a sua corrente de entrada máxima, aproximadamente 300 µA. Esta abordagem em tecnologia CMOS evita a utilização de um microssistema ótico de leitura de elevado custo, uma vez que torna possível a integração dos fotodíodos e respetiva eletrónica de leitura numa área de silício pequena (275 × 100 µm2 por fotodíodo e respetivo conversor). Foi avaliado o desempenho do microssistema implementado e dos filtros óticos fabricados usando fantomas (mais uma vez contendo hemoglobina, microesferas de polistireno, NADH e Carbostyril 124). Os resultados obtidos provaram a viabilidade do microssistema (incluindo o sistema de filtros óticos) para a extração de sinais de refletância difusa e de fluorescência. Foram notados alguns problemas na sensibilidade dos setups óticos implementados para as medições on-chip. No entanto, foram também propostas algumas soluções para os respetivos problemas, especificamente o uso futuro de díodos emissores de luz miniaturizados e a deposição direta dos filtros óticos no sistema de fotodeteção. Finalmente, a integração dos filtros óticos depositados diretamente em cima dos fotodíodos foi discutida e uma nova abordagem foi testada

Multimodal Multispectral Optical Endoscopic Imaging for Biomedical Applications

Optical imaging is an emerging field of clinical diagnostics that can address the growing medical need for early cancer detection and diagnosis. Various human cancers are amenable to better prognosis and patient survival if found and treated during early disease onset. Besides providing wide-field, macroscopic diagnostic information similar to existing clinical imaging techniques, optical imaging modalities have the added advantage of microscopic, high resolution cellular-level imaging from in vivo tissues in real time. This comprehensive imaging approach to cancer detection and the possibility of performing an ‘optical biopsy’ without tissue removal has led to growing interest in the field with numerous techniques under investigation. Three optical techniques are discussed in this thesis, namely multispectral fluorescence imaging (MFI), hyperspectral reflectance imaging (HRI) and fluorescence confocal endomicroscopy (FCE). MFI and HRI are novel endoscopic imaging-based extensions of single point detection techniques, such as laser induced fluorescence spectroscopy and diffuse reflectance spectroscopy. This results in the acquisition of spectral data in an intuitive imaging format that allows for quantitative evaluation of tissue disease states. We demonstrate MFI and HRI on fluorophores, tissue phantoms and ex vivo tissues and present the results as an RGB colour image for more intuitive assessment. This follows dimensionality reduction of the acquired spectral data with a fixed-reference isomap diagnostic algorithm to extract only the most meaningful data parameters. FCE is a probe-based point imaging technique offering confocal detection in vivo with almost histology-grade images. We perform FCE imaging on chemotherapy-treated in vitro human ovarian cancer cells, ex vivo human cancer tissues and photosensitiser-treated in vivo murine tumours to show the enhanced detection capabilities of the technique. Finally, the three modalities are applied in combination to demonstrate an optical viewfinder approach as a possible minimally-invasive imaging method for early cancer detection and diagnosis

Spectroscopy systems for the detection of gastrointestinal dysplasia

Programa doutoral em BioengenhariaThe detection of gastrointestinal (GI) dysplasia is essential to improve the patient’s survival rate. The diagnosis of this condition can be performed using spectroscopy techniques, such as fluorescence and diffuse-reflectance, which have the potential to provide morphological and biochemical information regarding normal and dysplastic tissue. Research prototypes currently used for those clinical spectroscopy techniques have associated a few drawbacks: they are costly, bulky, too sophisticated and they use optical fibers, which are usually related with low-collection efficiency. Moreover, these catheterbased instruments are invasive and very uncomfortable for the patient. In this context, the present work had the purpose to develop a miniaturized spectroscopy system, based on those two techniques, that features low-complexity and costeffectiveness. Furthermore, the integration of optical components in a single chip allows a high level of reliability. Such a system can be integrated in less-invasive devices (e.g., the endoscopic capsules), for an effective and comfortable detection of GI dysplastic lesions. The developed spectroscopy system core is based on thin-film optical filters and lowcost silicon photodiodes, used for the selection and detection of a few light wavelengths significant for the diagnosis of dysplasia. Thin-film optical filters, centered at specific wavelengths, were designed, fabricated and characterized after the successful demonstration that the use of only 16 spectral bands (within the 350 to 750 nm spectral range) enabled an accurate extraction of tissue information. The feasibility of using the fabricated filters to establish a quantitative spectroscopy diagnosis was proved with measurements on tissue phantoms. Additionally, an even more compact device is proposed for qualitative diagnosis using only two different spectral bands, centered at 420 nm for fluorescence measurements and at 540 nm for diffuse-reflectance measurements. This approach enabled the construction of a diagnostic algorithm for the identification of dysplastic tissues, with a sensitivity and specificity of 77.8% and 97.6%, respectively. This thesis work was also directed towards the development of a fluorescence and diffuse-reflectance spectroscopy imaging system for excised tissue margins assessment, which results from mucosal resections in the GI tract. In the developed system, wide area imaging is achieved by mechanically scanning an optical probe along the tissue surface, with variable spatial resolution. This medical device can provide real-time feedback regarding the resected mucosal margins, which represents a huge impact in intra-operative diagnosis. The clinical utility of the spectroscopy imaging prototype was successfully demonstrated using biological samples, i.e., different images containing reliable quantitative tissue information were obtained using analytical models. These images can be subsequently used to establish a diagnosis. This system may enable the reduction of the patient anxiety, avoiding the follow-up surgery, once a fast and real-time data analysis can be performed inside the operating room.A deteção de displasia gastrointestinal (GI) é essencial para melhorar a taxa de sobrevivência dos pacientes. O diagnóstico desta condição pode ser efetuado utilizando técnicas de espectroscopia, como a fluorescência e a reflectância difusa, que têm a capacidade de proporcionar informação morfológica e bioquímica acerca dos tecidos normais e dos tecidos com displasia. Os protótipos, ainda em investigação, actualmente utilizados em espetroscopia clínica e que implementam as técnicas mencionadas, têm associadas várias desvantagens, como por exemplo: têm elevado custo; são demasiado sofisticados e volumosos; e utilizam fibras óticas, que normalmente estão associadas a uma baixa eficiência na recolha de sinal. Para além disso, estes instrumentos baseiam-se em catéteres, o que os torna invasivos e, por isso, desconfortáveis para o paciente. Neste contexto, o presente trabalho teve como objetivo o desenvolvimento de um sistema baseado nas duas técnicas de espectroscopia acima referidas, que seja simples, de baixo custo e miniaturizado. A integração de componentes óticos num único chip possibilita a obtenção dispositivos óticos mais fiáveis e estáveis. Um sistema com estas características pode ser integrado em dispositivos de diagnóstico menos invasivos (por exemplo, nas cápsulas endoscópicas) para uma deteção eficaz e confortável das lesões displásticas do trato GI. O elemento principal deste sistema miniaturizado de espetroscopia consiste em filtros óticos baseados em filmes finos e fotodíodos de silício de baixo custo, ambos utilizados para a seleção e deteção de alguns comprimentos de onda importantes para o diagnóstico de displasia. Estes filtros óticos, centrados em comprimentos de onda específicos, foram desenhados, fabricados e caracterizados após ter sido demonstrado que a utilização de apenas 16 bandas espetrais (na gama espetral compreendida entre os 350 e os 750 nm) permitia uma extração correta da informação dos tecidos. A viabilidade de utilizar os filtros fabricados para o estabelecimento de um diagnóstico espetroscópico quantitativo foi também demonstrada com êxito através de medições realizadas em fantomas. Adicionalmente, e tendo em vista o desenvolvimento de um dispositivo ainda mais compacto, foi proposto um diagnóstico qualitativo, ou empírico, utilizando apenas duas bandas espetrais, centradas em 420 nm no caso das medições de fluorescência e em 540 nm no caso das medições de reflectância difusa. Esta abordagem permitiu a construção de um algoritmo de diagnóstico para a identificação de displasia com uma sensibilidade e uma especificidade de 77,8% e de 97,6%, respetivamente. Desenvolveu-se, igualmente, um sistema espetroscópico de imagem (envolvendo fluorescência e reflectância difusa) para a análise de margens de tecido excisado que resultam de ressecções da mucosa do tracto GI. Este sistema, através do varrimento mecânico de uma sonda de fibra ótica ao longo da superfície do tecido, com uma resolução espacial variável, possibilita a aquisição de imagens numa vasta área de tecido. Este dispositivo médico pode fornecer uma resposta em tempo real acerca das margens de tecido excisado, o que representa um enorme impacto no diagnóstico intra-operatório. A sua utilidade clínica foi demonstrada, com bons resultados, utilizando amostras biológicas, isto é, foi possível obter imagens contendo informação quantitativa fidedigna acerca dos tecidos, que pode ser subsequentemente utilizada para estabelecer um diagnóstico. Um sistema como este poderá reduzir a ansiedade dos pacientes e evitar a realização de novas cirurgias, uma vez que logo após a cirurgia, e ainda na sala de operações, pode efectuar-se uma análise aos tecidos.Fundação para a Ciência e a Tecnologia (FCT) - SFRH/BD/38978/2007MIT Portugal Progra

Depth-sensitive optical spectroscopy for noninvasive diagnosis of oral neoplasia

Oral cancer is the 11th most common cancer in the world. Cancers of the oral cavity and oropharynx account for more than 7,500 deaths each year in the United States alone. Major advances have been made in the management of oral cancer through the combined use of surgery, radiotherapy and chemotherapy, improving the quality of life for many patients; however, these advances have not led to a significant increase in survival rates, primarily because diagnosis often occurs at a late stage when treatment is more difficult and less successful. Accurate, objective, noninvasive methods for early diagnosis of oral neoplasia are needed. Here a method is presented to noninvasively evaluate oral lesions using depth-sensitive optical spectroscopy (DSOS). A ball lens coupled fiber-optic probe was developed to enable preferential targeting of different depth regions in the oral mucosa. Clinical studies of the diagnostic performance of DSOS in 157 subjects were carried out in collaboration with the University of Texas M. D. Anderson Cancer Center. An overall sensitivity of 90% and specificity of 89% were obtained for nonkeratinized oral tissue relative to histopathology. Based on these results a compact, portable version of the clinical DSOS device with real-time automated diagnostic capability was developed. The portable device was tested in 47 subjects and a sensitivity of 82% and specificity of 83% were obtained for nonkeratinized oral tissue. The diagnostic potential of multimodal platforms incorporating DSOS was explored through two pilot studies. A pilot study of DSOS in combination with widefield imaging was carried out in 29 oral cancer patients, resulting in a combined sensitivity of 94% and specificity of 69%. Widefield imaging and spectroscopy performed slightly better in combination than each method performed independently. A pilot study of DSOS in combination with the optical contrast agents 2-NBDG, EGF-Alexa 647, and proflavine was carried out in resected tissue specimens from 15 oral cancer patients. Improved contrast between neoplastic and healthy tissue was observed using 2-NBDG and EGF-Alexa 647

Development of deep learning methods for head and neck cancer detection in hyperspectral imaging and digital pathology for surgical guidance

Surgeons performing routine cancer resections utilize palpation and visual inspection, along with time-consuming microscopic tissue analysis, to ensure removal of cancer. Despite this, inadequate surgical cancer margins are reported for up to 10-20% of head and neck squamous cell carcinoma (SCC) operations. There exists a need for surgical guidance with optical imaging to ensure complete cancer resection in the operating room. The objective of this dissertation is to evaluate hyperspectral imaging (HSI) as a non-contact, label-free optical imaging modality to provide intraoperative diagnostic information. For comparison of different optical methods, autofluorescence, RGB composite images synthesized from HSI, and two fluorescent dyes are also acquired and investigated for head and neck cancer detection. A novel and comprehensive dataset was obtained of 585 excised tissue specimens from 204 patients undergoing routine head and neck cancer surgeries. The first aim was to use SCC tissue specimens to determine the potential of HSI for surgical guidance in the challenging task of head and neck SCC detection. It is hypothesized that HSI could reduce time and provide quantitative cancer predictions. State-of-the-art deep learning algorithms were developed for SCC detection in 102 patients and compared to other optical methods. HSI detected SCC with a median AUC score of 85%, and several anatomical locations demonstrated good SCC detection, such as the larynx, oropharynx, hypopharynx, and nasal cavity. To understand the ability of HSI for SCC detection, the most important spectral features were calculated and correlated with known cancer physiology signals, notably oxygenated and deoxygenated hemoglobin. The second aim was to evaluate HSI for tumor detection in thyroid and salivary glands, and RGB images were synthesized using the spectral response curves of the human eye for comparison. Using deep learning, HSI detected thyroid tumors with 86% average AUC score, which outperformed fluorescent dyes and autofluorescence, but HSI-synthesized RGB imagery performed with 90% AUC score. The last aim was to develop deep learning algorithms for head and neck cancer detection in hundreds of digitized histology slides. Slides containing SCC or thyroid carcinoma can be distinguished from normal slides with 94% and 99% AUC scores, respectively, and SCC and thyroid carcinoma can be localized within whole-slide images with 92% and 95% AUC scores, respectively. In conclusion, the outcomes of this thesis work demonstrate that HSI and deep learning methods could aid surgeons and pathologists in detecting head and neck cancers.Ph.D