Classification of brain tumours from MR spectra: the INTERPRET collaboration and its outcomes.

The INTERPRET project was a multicentre European collaboration, carried out from 2000 to 2002, which developed a decision-support system (DSS) for helping neuroradiologists with no experience of MRS to utilize spectroscopic data for the diagnosis and grading of human brain tumours. INTERPRET gathered a large collection of MR spectra of brain tumours and pseudo-tumoural lesions from seven centres. Consensus acquisition protocols, a standard processing pipeline and strict methods for quality control of the aquired data were put in place. Particular emphasis was placed on ensuring the diagnostic certainty of each case, for which all cases were evaluated by a clinical data validation committee. One outcome of the project is a database of 304 fully validated spectra from brain tumours, pseudotumoural lesions and normal brains, along with their associated images and clinical data, which remains available to the scientific and medical community. The second is the INTERPRET DSS, which has continued to be developed and clinically evaluated since the project ended. We also review here the results of the post-INTERPRET period. We evaluate the results of the studies with the INTERPRET database by other consortia or research groups. A summary of the clinical evaluations that have been performed on the post-INTERPRET DSS versions is also presented. Several have shown that diagnostic certainty can be improved for certain tumour types when the INTERPRET DSS is used in conjunction with conventional radiological image interpretation. About 30 papers concerned with the INTERPRET single-voxel dataset have so far been published. We discuss stengths and weaknesses of the DSS and the lessons learned. Finally we speculate on how the INTERPRET concept might be carried into the future.Funding from project MARESCAN (SAF2011-23870) from Ministerio de Economia y Competitividad in Spain. This work was also partially funded by CIBER-BBN, which is an initiative of the VI National R&D&i Plan 2008-2011, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. JRG acknowledges support from Cancer Research UK, the University of Cambridge and Hutchison Whampoa Ltd.This is the author accepted manuscript. The final version is available from Wiley via http://dx.doi.org/10.1002/nbm.343

Non-negative matrix factorisation methods for the spectral decomposition of MRS data from human brain tumours

<p>Abstract</p> <p>Background</p> <p><it>In-vivo </it>single voxel proton magnetic resonance spectroscopy (SV ¹H-MRS), coupled with supervised pattern recognition (PR) methods, has been widely used in clinical studies of discrimination of brain tumour types and follow-up of patients bearing abnormal brain masses. SV ¹H-MRS provides useful biochemical information about the metabolic state of tumours and can be performed at short (< 45 ms) or long (> 45 ms) echo time (TE), each with particular advantages. Short-TE spectra are more adequate for detecting lipids, while the long-TE provides a much flatter signal baseline in between peaks but also negative signals for metabolites such as lactate. Both, lipids and lactate, are respectively indicative of specific metabolic processes taking place. Ideally, the information provided by both TE should be of use for clinical purposes. In this study, we characterise the performance of a range of Non-negative Matrix Factorisation (NMF) methods in two respects: first, to derive sources correlated with the mean spectra of known tissue types (tumours and normal tissue); second, taking the best performing NMF method for source separation, we compare its accuracy for class assignment when using the mixing matrix directly as a basis for classification, as against using the method for dimensionality reduction (DR). For this, we used SV ¹H-MRS data with positive and negative peaks, from a widely tested SV ¹H-MRS human brain tumour database.</p> <p>Results</p> <p>The results reported in this paper reveal the advantage of using a recently described variant of NMF, namely Convex-NMF, as an unsupervised method of source extraction from SV¹H-MRS. Most of the sources extracted in our experiments closely correspond to the mean spectra of some of the analysed tumour types. This similarity allows accurate diagnostic predictions to be made both in fully unsupervised mode and using Convex-NMF as a DR step previous to standard supervised classification. The obtained results are comparable to, or more accurate than those obtained with supervised techniques.</p> <p>Conclusions</p> <p>The unsupervised properties of Convex-NMF place this approach one step ahead of classical label-requiring supervised methods for the discrimination of brain tumour types, as it accounts for their increasingly recognised molecular subtype heterogeneity. The application of Convex-NMF in computer assisted decision support systems is expected to facilitate further improvements in the uptake of MRS-derived information by clinicians.</p

Mass spectral imaging of clinical samples using deep learning

A better interpretation of tumour heterogeneity and variability is vital for the improvement of novel diagnostic techniques and personalized cancer treatments. Tumour tissue heterogeneity is characterized by biochemical heterogeneity, which can be investigated by unsupervised metabolomics. Mass Spectrometry Imaging (MSI) combined with Machine Learning techniques have generated increasing interest as analytical and diagnostic tools for the analysis of spatial molecular patterns in tissue samples. Considering the high complexity of data produced by the application of MSI, which can consist of many thousands of spectral peaks, statistical analysis and in particular machine learning and deep learning have been investigated as novel approaches to deduce the relationships between the measured molecular patterns and the local structural and biological properties of the tissues. Machine learning have historically been divided into two main categories: Supervised and Unsupervised learning. In MSI, supervised learning methods may be used to segment tissues into histologically relevant areas e.g. the classification of tissue regions in H&E (Haemotoxylin and Eosin) stained samples. Initial classification by an expert histopathologist, through visual inspection enables the development of univariate or multivariate models, based on tissue regions that have significantly up/down-regulated ions. However, complex data may result in underdetermined models, and alternative methods that can cope with high dimensionality and noisy data are required. Here, we describe, apply, and test a novel diagnostic procedure built using a combination of MSI and deep learning with the objective of delineating and identifying biochemical differences between cancerous and non-cancerous tissue in metastatic liver cancer and epithelial ovarian cancer. The workflow investigates the robustness of single (1D) to multidimensional (3D) tumour analyses and also highlights possible biomarkers which are not accessible from classical visual analysis of the H&E images. The identification of key molecular markers may provide a deeper understanding of tumour heterogeneity and potential targets for intervention.Open Acces

Modelling FTIR spectral sata with Type-I and Type-II fuzzy sets for breast cancer grading

Breast cancer is one of the most frequently occurring cancers amongst women throughout the world. After the diagnosis of the disease, monitoring its progression is important in predicting the chances of long term survival of patients. The Nottingham Prognostic Index (NPI) is one of the most common indices used to categorise the patients into different groups depending upon the severity of the disease. One of the key factors of this index is cancer grade which is determined by pathologists who examine cell samples under a microscope. This manual method has a higher chance of false classification and may lead to incorrect treatment of patients. There is a need to develop automated methods that employ advanced computational methods to help pathologists in making a decision regarding the classification of breast cancer grade. Fourier transform infra-red spectroscopy (FTIR) is one of the relatively new techniques that has been used for diagnosis of various cancer types with advanced computational methods in the literature. In this thesis we examine the use of advanced fuzzy methods with the FTIR spectral data sets to develop a model prototype that can help clinicians with breast cancer grading. Initial work is focussed on using the commonly used clustering algorithms k-means and fuzzy c-means with principal component analysis on different cancer spectral data sets to explore the complexities within them. After that, a novel model based on Type-II fuzzy logic is developed for use on a complex breast cancer FTIR spectral data set that can help clinicians classify breast cancer grades. The data set used for the purpose consists of multiple cases of each grade. We consider two types of uncertainty, one within the spectra of a single case of a grade (intra -case) and other when comparing it with other cases of same grade (inter-case). Features have been extracted in terms of interval data from various peaks and troughs. The interval data from the features has been used to create Type-I fuzzy sets for each case. After that the Type-I fuzzy sets are combined to create zSlices based General Type-II fuzzy sets for each feature for each grade. The created benchmark fuzzy sets are then used as prototypes for classification of unseen spectral data. Type-I fuzzy sets are created for unseen spectral data and then compared against the benchmark prototype Type-II fuzzy sets for each grade using a similarity measure. The best match based on the calculated similarity scores is assigned as the resultant grade. The novel model is tested on an independent spectral data set of oral cancer patients. Results indicate that the model was able to successfully construct prototype fuzzy sets for the data set, and provide in-depth information regarding the complexities of the data set as well as helping in classification of the data

Machine learning strategies for diagnostic imaging support on histopathology and optical coherence tomography

Tesis por compendio[ES] Esta tesis presenta soluciones de vanguardia basadas en algoritmos de computer vision (CV) y machine learning (ML) para ayudar a los expertos en el diagnóstico clínico. Se centra en dos áreas relevantes en el campo de la imagen médica: la patología digital y la oftalmología. Este trabajo propone diferentes paradigmas de machine learning y deep learning para abordar diversos escenarios de supervisión en el estudio del cáncer de próstata, el cáncer de vejiga y el glaucoma. En particular, se consideran métodos supervisados convencionales para segmentar y clasificar estructuras específicas de la próstata en imágenes histológicas digitalizadas. Para el reconocimiento de patrones específicos de la vejiga, se llevan a cabo enfoques totalmente no supervisados basados en técnicas de deep-clustering. Con respecto a la detección del glaucoma, se aplican algoritmos de memoria a corto plazo (LSTMs) que permiten llevar a cabo un aprendizaje recurrente a partir de volúmenes de tomografía por coherencia óptica en el dominio espectral (SD-OCT). Finalmente, se propone el uso de redes neuronales prototípicas (PNN) en un marco de few-shot learning para determinar el nivel de gravedad del glaucoma a partir de imágenes OCT circumpapilares. Los métodos de inteligencia artificial (IA) que se detallan en esta tesis proporcionan una valiosa herramienta de ayuda al diagnóstico por imagen, ya sea para el diagnóstico histológico del cáncer de próstata y vejiga o para la evaluación del glaucoma a partir de datos de OCT.[CA] Aquesta tesi presenta solucions d'avantguarda basades en algorismes de *computer *vision (CV) i *machine *learning (ML) per a ajudar als experts en el diagnòstic clínic. Se centra en dues àrees rellevants en el camp de la imatge mèdica: la patologia digital i l'oftalmologia. Aquest treball proposa diferents paradigmes de *machine *learning i *deep *learning per a abordar diversos escenaris de supervisió en l'estudi del càncer de pròstata, el càncer de bufeta i el glaucoma. En particular, es consideren mètodes supervisats convencionals per a segmentar i classificar estructures específiques de la pròstata en imatges histològiques digitalitzades. Per al reconeixement de patrons específics de la bufeta, es duen a terme enfocaments totalment no supervisats basats en tècniques de *deep-*clustering. Respecte a la detecció del glaucoma, s'apliquen algorismes de memòria a curt termini (*LSTMs) que permeten dur a terme un aprenentatge recurrent a partir de volums de tomografia per coherència òptica en el domini espectral (SD-*OCT). Finalment, es proposa l'ús de xarxes neuronals *prototípicas (*PNN) en un marc de *few-*shot *learning per a determinar el nivell de gravetat del glaucoma a partir d'imatges *OCT *circumpapilares. Els mètodes d'intel·ligència artificial (*IA) que es detallen en aquesta tesi proporcionen una valuosa eina d'ajuda al diagnòstic per imatge, ja siga per al diagnòstic histològic del càncer de pròstata i bufeta o per a l'avaluació del glaucoma a partir de dades d'OCT.[EN] This thesis presents cutting-edge solutions based on computer vision (CV) and machine learning (ML) algorithms to assist experts in clinical diagnosis. It focuses on two relevant areas at the forefront of medical imaging: digital pathology and ophthalmology. This work proposes different machine learning and deep learning paradigms to address various supervisory scenarios in the study of prostate cancer, bladder cancer and glaucoma. In particular, conventional supervised methods are considered for segmenting and classifying prostate-specific structures in digitised histological images. For bladder-specific pattern recognition, fully unsupervised approaches based on deep-clustering techniques are carried out. Regarding glaucoma detection, long-short term memory algorithms (LSTMs) are applied to perform recurrent learning from spectral-domain optical coherence tomography (SD-OCT) volumes. Finally, the use of prototypical neural networks (PNNs) in a few-shot learning framework is proposed to determine the severity level of glaucoma from circumpapillary OCT images. The artificial intelligence (AI) methods detailed in this thesis provide a valuable tool to aid diagnostic imaging, whether for the histological diagnosis of prostate and bladder cancer or glaucoma assessment from OCT data.García Pardo, JG. (2022). Machine learning strategies for diagnostic imaging support on histopathology and optical coherence tomography [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/182400Compendi

Modelling FTIR spectral sata with Type-I and Type-II fuzzy sets for breast cancer grading

Breast cancer is one of the most frequently occurring cancers amongst women throughout the world. After the diagnosis of the disease, monitoring its progression is important in predicting the chances of long term survival of patients. The Nottingham Prognostic Index (NPI) is one of the most common indices used to categorise the patients into different groups depending upon the severity of the disease. One of the key factors of this index is cancer grade which is determined by pathologists who examine cell samples under a microscope. This manual method has a higher chance of false classification and may lead to incorrect treatment of patients. There is a need to develop automated methods that employ advanced computational methods to help pathologists in making a decision regarding the classification of breast cancer grade. Fourier transform infra-red spectroscopy (FTIR) is one of the relatively new techniques that has been used for diagnosis of various cancer types with advanced computational methods in the literature. In this thesis we examine the use of advanced fuzzy methods with the FTIR spectral data sets to develop a model prototype that can help clinicians with breast cancer grading. Initial work is focussed on using the commonly used clustering algorithms k-means and fuzzy c-means with principal component analysis on different cancer spectral data sets to explore the complexities within them. After that, a novel model based on Type-II fuzzy logic is developed for use on a complex breast cancer FTIR spectral data set that can help clinicians classify breast cancer grades. The data set used for the purpose consists of multiple cases of each grade. We consider two types of uncertainty, one within the spectra of a single case of a grade (intra -case) and other when comparing it with other cases of same grade (inter-case). Features have been extracted in terms of interval data from various peaks and troughs. The interval data from the features has been used to create Type-I fuzzy sets for each case. After that the Type-I fuzzy sets are combined to create zSlices based General Type-II fuzzy sets for each feature for each grade. The created benchmark fuzzy sets are then used as prototypes for classification of unseen spectral data. Type-I fuzzy sets are created for unseen spectral data and then compared against the benchmark prototype Type-II fuzzy sets for each grade using a similarity measure. The best match based on the calculated similarity scores is assigned as the resultant grade. The novel model is tested on an independent spectral data set of oral cancer patients. Results indicate that the model was able to successfully construct prototype fuzzy sets for the data set, and provide in-depth information regarding the complexities of the data set as well as helping in classification of the data

The real-time molecular characterisation of human brain tumours during surgery using Rapid Evaporative Ionization Mass Spectrometry [REIMS] and Raman spectroscopy: a platform for precision medicine in neurosurgery

Aim: To investigate new methods for the chemical detection of tumour tissue during neurosurgery. Rationale: Surgeons operating on brain tumours currently lack the ability to directly and immediately assess the presence of tumour tissue to help guide resection. Through developing a first in human application of new technology we hope to demonstrate the proof of concept that chemical detection of tumour tissue is possible. It will be further demonstrated that information can be obtained to potentially aid treatment decisions. This new technology could, therefore, become a platform for more effective surgery and introducing precision medicine to Neurosurgery. Methods: Molecular analysis was performed using Raman spectroscopy and Rapid Evaporative Ionization Mass Spectrometry (REIMS). These systems were first developed for use in brain surgery. A single centre prospective observational study of both modalities was designed involving a total of 75 patients undergoing craniotomy and resection of a range of brain tumours. A neuronavigation system was used to register spectral readings in 3D space. Precise intraoperative readings from different tumour zones were taken and compared to matched core biopsy samples verified by routine histopathology. Results: Multivariate statistics including PCA/LDA analysis was used to analyse the spectra obtained and compare these to the histological data. The systems identified normal versus tumour tissue, tumour grade, tumour type, tumour density and tissue status of key markers of gliomagenesis. Conclusions: The work in this thesis provides proof of concept that useful real time intraoperative spectroscopy is possible. It can integrate well with the current operating room setup to provide key information which could potentially enhance surgical safety and effectiveness in increasing extent of resection. The ability to group tissue samples with respect to genomic data opens up the possibility of using this information during surgery to speed up treatment, escalate/deescalate surgery in specific phenotypic groups to introduce precision medicine to Neurosurgery.Open Acces

Multivariate methods for interpretable analysis of magnetic resonance spectroscopy data in brain tumour diagnosis

Malignant tumours of the brain represent one of the most difficult to treat types of cancer due to the sensitive organ they affect. Clinical management of the pathology becomes even more intricate as the tumour mass increases due to proliferation, suggesting that an early and accurate diagnosis is vital for preventing it from its normal course of development. The standard clinical practise for diagnosis includes invasive techniques that might be harmful for the patient, a fact that has fostered intensive research towards the discovery of alternative non-invasive brain tissue measurement methods, such as nuclear magnetic resonance. One of its variants, magnetic resonance imaging, is already used in a regular basis to locate and bound the brain tumour; but a complementary variant, magnetic resonance spectroscopy, despite its higher spatial resolution and its capability to identify biochemical metabolites that might become biomarkers of tumour within a delimited area, lags behind in terms of clinical use, mainly due to its difficult interpretability. The interpretation of magnetic resonance spectra corresponding to brain tissue thus becomes an interesting field of research for automated methods of knowledge extraction such as machine learning, always understanding its secondary role behind human expert medical decision making. The current thesis aims at contributing to the state of the art in this domain by providing novel techniques for assistance of radiology experts, focusing on complex problems and delivering interpretable solutions. In this respect, an ensemble learning technique to accurately discriminate amongst the most aggressive brain tumours, namely glioblastomas and metastases, has been designed; moreover, a strategy to increase the stability of biomarker identification in the spectra by means of instance weighting is provided. From a different analytical perspective, a tool based on signal source separation, guided by tumour type-specific information has been developed to assess the existence of different tissues in the tumoural mass, quantifying their influence in the vicinity of tumoural areas. This development has led to the derivation of a probabilistic interpretation of some source separation techniques, which provide support for uncertainty handling and strategies for the estimation of the most accurate number of differentiated tissues within the analysed tumour volumes. The provided strategies should assist human experts through the use of automated decision support tools and by tackling interpretability and accuracy from different anglesEls tumors cerebrals malignes representen un dels tipus de càncer més difícils de tractar degut a la sensibilitat de l’òrgan que afecten. La gestió clínica de la patologia esdevé encara més complexa quan la massa tumoral s'incrementa degut a la proliferació incontrolada de cèl·lules; suggerint que una diagnosis precoç i acurada és vital per prevenir el curs natural de desenvolupament. La pràctica clínica estàndard per a la diagnosis inclou la utilització de tècniques invasives que poden arribar a ser molt perjudicials per al pacient, factor que ha fomentat la recerca intensiva cap al descobriment de mètodes alternatius de mesurament dels teixits del cervell, tals com la ressonància magnètica nuclear. Una de les seves variants, la imatge de ressonància magnètica, ja s'està actualment utilitzant de forma regular per localitzar i delimitar el tumor. Així mateix, una variant complementària, la espectroscòpia de ressonància magnètica, malgrat la seva alta resolució espacial i la seva capacitat d'identificar metabòlits bioquímics que poden esdevenir biomarcadors de tumor en una àrea delimitada, està molt per darrera en termes d'ús clínic, principalment per la seva difícil interpretació. Per aquest motiu, la interpretació dels espectres de ressonància magnètica corresponents a teixits del cervell esdevé un interessant camp de recerca en mètodes automàtics d'extracció de coneixement tals com l'aprenentatge automàtic, sempre entesos com a una eina d'ajuda per a la presa de decisions per part d'un metge expert humà. La tesis actual té com a propòsit la contribució a l'estat de l'art en aquest camp mitjançant l'aportació de noves tècniques per a l'assistència d'experts radiòlegs, centrades en problemes complexes i proporcionant solucions interpretables. En aquest sentit, s'ha dissenyat una tècnica basada en comitè d'experts per a una discriminació acurada dels diferents tipus de tumors cerebrals agressius, anomenats glioblastomes i metàstasis; a més, es proporciona una estratègia per a incrementar l'estabilitat en la identificació de biomarcadors presents en un espectre mitjançant una ponderació d'instàncies. Des d'una perspectiva analítica diferent, s'ha desenvolupat una eina basada en la separació de fonts, guiada per informació específica de tipus de tumor per a avaluar l'existència de diferents tipus de teixits existents en una massa tumoral, quantificant-ne la seva influència a les regions tumorals veïnes. Aquest desenvolupament ha portat cap a la derivació d'una interpretació probabilística d'algunes d'aquestes tècniques de separació de fonts, proporcionant suport per a la gestió de la incertesa i estratègies d'estimació del nombre més acurat de teixits diferenciats en cada un dels volums tumorals analitzats. Les estratègies proporcionades haurien d'assistir els experts humans en l'ús d'eines automatitzades de suport a la decisió, donada la interpretabilitat i precisió que presenten des de diferents angles

Texture Analysis Platform for Imaging Biomarker Research

abstract: The rate of progress in improving survival of patients with solid tumors is slow due to late stage diagnosis and poor tumor characterization processes that fail to effectively reflect the nature of tumor before treatment or the subsequent change in its dynamics because of treatment. Further advancement of targeted therapies relies on advancements in biomarker research. In the context of solid tumors, bio-specimen samples such as biopsies serve as the main source of biomarkers used in the treatment and monitoring of cancer, even though biopsy samples are susceptible to sampling error and more importantly, are local and offer a narrow temporal scope. Because of its established role in cancer care and its non-invasive nature imaging offers the potential to complement the findings of cancer biology. Over the past decade, a compelling body of literature has emerged suggesting a more pivotal role for imaging in the diagnosis, prognosis, and monitoring of diseases. These advances have facilitated the rise of an emerging practice known as Radiomics: the extraction and analysis of large numbers of quantitative features from medical images to improve disease characterization and prediction of outcome. It has been suggested that radiomics can contribute to biomarker discovery by detecting imaging traits that are complementary or interchangeable with other markers. This thesis seeks further advancement of imaging biomarker discovery. This research unfolds over two aims: I) developing a comprehensive methodological pipeline for converting diagnostic imaging data into mineable sources of information, and II) investigating the utility of imaging data in clinical diagnostic applications. Four validation studies were conducted using the radiomics pipeline developed in aim I. These studies had the following goals: (1 distinguishing between benign and malignant head and neck lesions (2) differentiating benign and malignant breast cancers, (3) predicting the status of Human Papillomavirus in head and neck cancers, and (4) predicting neuropsychological performances as they relate to Alzheimer’s disease progression. The long-term objective of this thesis is to improve patient outcome and survival by facilitating incorporation of routine care imaging data into decision making processes.Dissertation/ThesisDoctoral Dissertation Biomedical Informatics 201

Multimodal FTIR Microscopy-guided Acquisition and Interpretation of MALDI Mass Spectrometry Imaging Data

Multimodale klinische Bildgebung stellt eine der bedeutendsten Entwicklung der letzten Jahrzehnte dar. Neben der Kombination komplementärer in vivo Sensoren in beispielsweise PET-MRI oder SPECT-CT sind auch ex vivo Analyseverfahren, welche eine genauere Beschreibung der Probe ermöglichen, in den Bereich der (prä)klinischen Diagnostik vorgedrungen. Eine der vielversprechendsten Techniken in diesem Zusammenhang stellt die bildgebende Massenspektrometrie dar, welche die Verteilungsmuster hunderter Biomoleküle oder Pharmazeutika semi-quantitativ erfasst. Dabei kommt das Verfahren ohne die Verwendung von markierten Substanzen aus und erlaubt eine höhere räumliche und spektrale Auflösung im Vergleich zu in vivo Sensoren. Allerdings unterliegt die Technik auch einigen wesentlichen Einschränkungen, da die Datenakquisition besonders bei der Verwendung von ultrahochauflösenden FTICR-Detektoren sehr langsam erfolgt. Die niedrige Durchsatzleistung und damit verbundene unhandliche Datenmenge erschwert somit die Analyse größerer Patientenkohorten, wodurch ein Bedarf an multimodalen Lösungsansätzen besteht. Ein geeignetes Verfahren in dieser Hinsicht stellt die Schwingungsspektroskopie (bsp. Infrarotspektroskopie) dar, welche räumliche Details vergleichsweise schnell erfasst; dabei allerdings keine Rückschlüsse auf die Verteilung bestimmter chemischer Substanzen ermöglicht. Im Rahmen der vorliegenden Arbeit wurde ein MATLAB-gestütztes Verfahren zur multimodalen Akquirierung von Infrarotspektroskopie- und Massenspektrometrie-Daten entwickelt und bewertet. Dabei werden räumliche Strukturen und Zellpopulationen innerhalb von Geweben mittels FTIR-basierter Clusteranalyse segmentiert. Anschließend kann die chemische Zusammensetzung einzelner Segmente zielgerichtet akquiriert und verglichen werden. Das entwickelte Verfahren funktioniert dabei unabhängig von konventioneller histopathologischer Gewebeannotation. Ein wichtiger Faktor bei Mittelinfrarot- und Massenspektrometrie-Messungen auf Gewebe stellt die Zusammensetzung der verwendeten Objektträger-Beschichtung dar. Für die Bewertung der erhaltenen Spektren und der damit verbundenen Bildsegmentierung wurden deshalb Experimente auf Indiumzinnoxid, Silberzinnoxid und Gold durchgeführt und verglichen. Dabei konnte gezeigt werden, dass Infrarot- und Massenspektrometrie-Bilder von der gleichen Probe auf Gold mit hoher Qualität aufgenommen werden können. Weiterhin konnte gezeigt werden, dass durch einfache Infrarotsegmentierung eine Identifizierung relevanter morphologischer Gehirnstrukturen möglich ist. Die erzielte räumliche Präzision und Auflösung der Infrarot-Segmente stellt dabei einen deutlichen Mehrwert gegenüber der direkten Segmentierung von Massenspektrometriebildern dar. Darüber hinaus können Infrarotsegmente bereits vor der eigentlichen MS-Messung generiert werden. Nach erfolgter Methodenentwicklung und Validierung konnte diese auf verschiedene diagnostische Studien angewendet werden. In einem ersten Anwendungsbeispiel konnten in Mäuse xenotransplantierte humane Glioblastomzellen mit erhöhter Präzision visualisiert werden. Darüber hinaus wurde eine im korrespondierenden H&E-Bild unauffällige, den Tumor-umschließende Struktur identifiziert. Durch den erfolgreichen Transfer der Infrarotsegmente in das Koordinatensystem von nachfolgend gemessenen MS-Bildern, konnten spezifische Markersignaturen automatisch extrahiert werden. Im Zuge dessen konnte die Authentizität Tumorstruktur sowie der zweiten Tumor-assoziierten Struktur durch spezifische Massen bekräftigt werden. In einer weiteren Studie, wurde die entwickelte Methode für das automatische Screening von Markersignaturen in Niemann-Pick Typ C1 ähnlichen murinen Kleinhirnschnitten getestet. Dabei konnten regionsspezifische, im Gesamtdatensatz insignifikante Änderungen in der Lipidzusammensetzung automatisiert uns Annotations-unabhängig erfasst werden. In einer weiteren Infrarotspektroskopie-Studie an 89 kryokonservierten GIST Schnitten von 27 Patienten konnte eine schnelle und simultane Segmentierung aller Gewebeproben exemplarisch gezeigt werden. Dabei wurden farbkodierte Bilder aller Proben generiert, in denen gleiche Farben für eine spektrale Ähnlichkeit stehen. Durch den Abgleich der erhaltenen Farbcodes mit histopathologisch annotierten Folgeschnitten konnten zwei der fünf dargestellten Farbgruppen mit dem Auftreten von Tumorzellen assoziiert werden. Die anderen Gruppen repräsentierten Fibrosen, Nekrosen und weitere nicht-tumoröse Gewebeanteile. Abschließend wurde die Struktur-gerichtete Akquisition von ultrahochauflösenden FTICR-MS Bildern gezeigt, welche auf Basis von Mittelinfrarotbildern der identischen Gewebeprobe abgeleitet wurden. Indem die zeitaufwändige MS-Messung ausschließlich auf kleinere Strukturen von Interesse (wie beispielsweise die Körnerzell-Schicht der Cornu Ammonis) gerichtet wurde, konnte eine Zeit- und Datenersparnis von bis zu 97.8% gegenüber der vollständigen Messung erreicht werden. Damit ist ein großer Schritt hin zur Implementierung von ultrahochauflösender Massenspektrometrie im klinischen Umfeld erfolgt