[pt] Mais do que nunca, um dos desafios para o design de interação hoje é o desenvolvimento de sistemas que atendam às necessidades e expectativas de pessoas de diferentes origens culturais e sociais. A perspectiva mais amplamente adotada é a internacionalização-localização. Internacionalização é o processo de separação do código do núcleo funcional das especificidades da interface do sistema (por exemplo, o idioma, as medidas, etc.). Com a localização, a interface é customizada para um determinado público (através da tradução do idioma, dos marcadores culturais e até mesmo de características técnicas, por exemplo). Internacionalização e localização têm como resultado esconder ou neutralizar diferenças culturais entre as comunidades de usuários e contextos de uso distintos. Estamos, no entanto, interessados nas situações onde a intenção de design é praticamente oposta: explorar a diversidade cultural. Esta tese oferece conhecimento novo para ajudar designers de IHC a comunicarem sua intenção de design quando querem promover o contato dos usuários com a diversidade cultural. São apresentadas cinco metáforas de perspectivas culturais (CVM) para apoiar o raciocínio e a tomada de decisão sobre dimensões da experiência intercultural. As metáforas derivam de estudos empíricos aplicando Engenharia Semiótica para analisar e re-projetar interfaces de sistemas multiculturais. A fim de investigar se e como estas metáforas apoiam os profissionais de IHC em tempo de design e de avaliação, realizamos um estudo de caso para avaliar como as CVM podem ser usadas em atividades de re-projeto e avaliação. Descobrimos que as CVM desempenham um papel importante nos estágios iniciais do re-projeto e efetivamente ajudam os designers a raciocinar sobre possíveis experiências interculturais que poderão acontecer em tempo de interação. Além disso, as CVM fornecem uma rica grade epistêmica, onde a consistência das escolhas de projeto se destaca mais claramente. / [en] More than ever before, today one of the challenges for interaction design is the development of systems aiming to attend to the needs and expectations of people with different cultural and social backgrounds. The most widely used perspective in cross-cultural design is internationalization-localization. Internationalization is the process of separating the core functionality code from system s interface specifics (e.g. text language, measures, etc.). With localization, the interface is customized for a particular audience (through language translation, cultural markers and even technical features, for instance). The result of internationalization and localization is to conceal or neutralize cultural differences among different user communities and contexts of use. We are, however, interested in another situation: one where the design intent is virtually the opposite: to expose and explore cultural diversity. This is the case, for instance, when the purpose of the designed system is to stimulate users to make contact with a foreign culture. This thesis provides new knowledge to help HCI designers communicate their intent when they want to promote the users contact with cultural diversity. We present five cultural viewpoint metaphors (CVM) to support reasoning and decision-making about intercultural experience dimensions. The metaphors derive from empirical studies applying Semiotic Engineering to analyze and re-design cross-cultural systems interfaces. In order to investigate if and how CVM actually support HCI professionals/practitioners at design and evaluation time, we carried out an extensive case study to assess how CVM can be used in design and evaluation activities. We found that CVM played an important role in early design stages, helping designers to reason effectively about intercultural experiences while determining which cultural perspective they want to adopt. Furthermore, CVM features provided a rich epistemic grid where the consistency of design choices stands out more clearly.

abstract: Eosinophils are innate immune cells that are most commonly associated with parasite infection and allergic responses. Recent studies, though, have identified eosinophils as cells with diverse effector functions at baseline and in disease. Eosinophils in specific tissue immune environments are proposed to promote unique and specific effector functions, suggesting these cells have the capacity to differentiate into unique subtypes. The studies here focus on defining these subtypes using functional, molecular, and genetic analysis as well as using novel techniques to image these subtypes in situ. To characterized these subtypes, an in vitro cytokine induced type 1 (E1) and type 2 (E2) eosinophil model was developed that display features and functions of eosinophils found in vivo. For example, E1 eosinophils secrete type 1 mediators (e.g., IL-12, CXCL9 and CXCL10), express iNOS and express increased levels of the surface molecules PDL1 and MHC-I. Conversely, E2 eosinophils release type 2 mediators (e.g., IL4, IL13, CCL17, and CCL22), degranulate and express increased surface molecules CD11b, ST2 and Siglec-F. Completion of differential expression analysis of RNAseq on these subtypes revealed 500 and 655 unique genes were upregulated in E1 and E2 eosinophils, respectively. Functional enrichment studies showed interferon regulatory factor (IRF) transcription factors were uniquely regulated in both mouse and human E1 and E2 eosinophils. These subtypes are sensitive to their environment, modulating their IRF and cell surface expression when stimulated with opposing cytokines, suggesting plasticity. To identify and study these subtypes in situ, chromogenic and fluorescent eosinophil-specific immunostaining protocols were developed. Methods were created and optimized, here, to identify eosinophils by their granule proteins in formalin fixed mouse tissues. Yet, eosinophil-specific antibodies alone are not enough to identify and study the complex interactions eosinophil subtypes perform within a tissue. Therefore, as part of this thesis, a novel highly-multiplexed immunohistochemistry technique was developed utilizing cleavable linkers to address these concerns. This technique is capable of analyzing up to 22 markers within a single biopsy with single-cell resolution. With this approach, eosinophil subtypes can be studied in situ in routine patient biopsies. / Dissertation/Thesis / Doctoral Dissertation Biochemistry 2020

[pt] É fato que a diversidade cultural tornou-se um novo desafio na Interação Humano-Computador. Atualmente, os usuários podem navegar por quase todos os lugares da Web, sem fronteiras nacionais e/ou culturais, fazendo contato intencional ou não com elementos de culturas estrangeiras (idiomas, práticas e etc.). A Web tornou-se, portanto, um local privilegiado para encontros interculturais, i.e., um lugar onde os usuários têm a oportunidade de entrar em contato com a diversidade cultural diretamente (ao interagir com outros usuários através de redes sociais, por exemplo) ou indiretamente (quando interagem com aplicações que carregam traços culturais estrangeiros). Este cenário indica a necessidade de uma investigação sobre como a metacomunicação (comunicação sobre a comunicação) intercultural dos designers para os usuários é percebida pelos usuários. Com isto objetivamos entender como os usuários percebem as oportunidades de estabelecer contato com a diversidade cultural ao interagirem com aplicações multiculturais e como essas percepções podem contribuir em atividades de Avaliação de IHC de tais sistemas. Então, a fim de investigar se e como os usuários expressam as suas percepções e reações sobre as oportunidades promovidas pelos encontros interculturais indiretos, realizamos estudos empíricos onde oferecemos um vocabulário específico (Metáforas de Perspectivas Culturais). Realizamos também outros estudos sem oferecer nenhum vocabulário, i.e., deixamos os usuários falarem livremente sobre as oportunidades de fazer contato com a diversidade cultural na Interação Humano-Computador. Esses estudos foram realizados no contexto de duas aplicações multiculturais (uma do domínio linguístico e outra de um domínio não linguístico). Entre os resultados obtidos, destaca-se o potencial do vocabulário específico no ciclo de design de interação de sistemas multiculturais, revelando que as Metáforas de Perspectivas Culturais são uma ferramenta de apoio promissora para as práticas de design participativo, i.e., um meio de expressão e comunicação para os usuários qualificarem suas experiências de interação reais ou potenciais. / [en] It is fact that cultural diversity has become a new challenge in Human-Computer Interaction. Today users can navigate almost anywhere on the Web, with no cultural and/or national boundaries, having intentional or unintentional contact with foreign cultural elements (languages, practices etc.). Therefore, the Web became a privileged place for intercultural encounters, i.e., a place where users have the opportunity to be in contact with cultural diversity directly (when interacting with other users using social networks, for example) or indirectly (when interacting with applications, which have foreign cultural features). This scenario indicates the need for an investigation about how the intercultural metacommunication (communication about communication) of designers to users is noticed by users. Hereupon, we intend to understand how users perceive the opportunities to make contact with cultural diversity when interacting with cross-cultural applications and how these perceptions can contribute in the HCI Evaluation activities of these systems. Thus, in order to investigate whether and how users express their perceptions and reactions over opportunities promoted by indirect intercultural encounters, we conducted empirical studies in which we offered a specific vocabulary (Cultural Viewpoint Metaphors). We also conducted other studies without offering any vocabulary, that is, we let users speak freely about the opportunities to make contact with cultural diversity in Human-Computer Interaction. These studies were conducted in the context of two cross-cultural applications (one in a linguistic domain and one in a non-linguistic domain). Among the results obtained, it was highlighted the potential of the specific vocabulary in the interaction design cycle of cross-cultural systems, revealing that Cultural Viewpoint Metaphors are a promising supporting tool for participatory design practices, i.e., a medium of expression and communication for users to qualify their real or potential interaction experiences.

Made available in DSpace on 2016-06-02T19:05:42Z (GMT). No. of bitstreams: 1 2997.pdf: 4158163 bytes, checksum: 540ba66fcb2c536c7a72d8add391f0b3 (MD5) Previous issue date: 2010-05-12 / Financiadora de Estudos e Projetos / The software reengineering becomes important because of the need that organizations have in adjusting to new trends, technologies and user requirements. The term organization should be understood like universities or companies that develop software for use by a large number of people. Thinking about the adequacy of users requirements, is presented in this work the reengineering process of a software called Cognitor, which is a tool designed to support teachers in the process of creating electronic teaching materials. The need to reengineer Cognitor was perceived through a case study where several changes were pointed by users. Some of these changes are: text editor improvement, preview of the images that are inserted in the content pages, feedback to the users, among others. During the reengineering of this software, it was formalized a software reengineering process model, user-centered, for the conversion of desktop applications in RIAs (Rich Internet Application), called UC-RIA (User Centered Rich Internet Application). The process model was named as UC-RIA due to the participation of the potential users during the application s reengineering process, because they were involved in prototyping and in validation of the graphical interfaces of the new version. The results of this study show the capability of the proposed software reengineering model to be used as a support in organizations for the reengineering of their software, mainly because it inserts the users in the reengineering process during the application s Prototyping phase, bringing software to users real needs. / A reengenharia de software se faz importante devido à necessidade que as organizações têm em se adequar às novas tendências, tecnologias e exigências dos usuários. Inclui-se ao termo organizações , empresas ou universidades que desenvolvem softwares para serem utilizados por um grande número de pessoas. Pensando na adequação das exigências dos usuários, este trabalho apresenta a reengenharia de um software chamado Cognitor, que é uma ferramenta criada para apoiar os educadores no processo de criação de material didático eletrônico. A percepção da necessidade da reengenharia do Cognitor se deu através de um estudo de caso onde foram relatadas várias alterações que o software deveria sofrer, tais como: melhoria no editor de texto, pré-visualização das imagens que são inseridas nas páginas de conteúdo, feedback ao usuário, entre outras. Durante a reengenharia desse software, consequentemente, foi formalizado um modelo de processo de reengenharia de software, centrado no usuário, para a conversão de aplicações desktop em RIAs (Rich Internet Application), denominado UC-RIA (User Centered Rich Internet Application). Foi dado o nome de UC-RIA ao modelo de processo devido à participação dos potenciais usuários da aplicação durante o processo de reengenharia, pois estes estiveram envolvidos tanto na prototipação, quanto na validação das interfaces gráficas da nova versão. Os resultados obtidos neste trabalho mostram a potencialidade do modelo de reengenharia de software UC-RIA em ser utilizado como apoio às organizações durante a reengenharia de seus softwares, principalmente por inserir os usuários no processo de reengenharia durante a fase de Prototipação da aplicação, aproximando os softwares às reais necessidades dos usuários.

Bladder cancer is a potentially curable malignancy; however in regards to the state of current therapy regimens, a plateau has been reached in both the non-muscle and muscle invasive types. To obtain effective treatment, and consequently a decreased mortality, it has become imperative to test and understand aspects affecting therapy response. The aim of this thesis is to illustrate a better understanding of clinical factors affecting therapy response using new drug combinations and new tumor markers alongside established risk criteria. In Paper I we reported the 5 year follow up from a multicenter, prospectively randomized study and we evaluated the 5-year outcomes of BCG alone compared to a combination of epirubicin and interferon-a2b in the treatment of patients with T1 bladder cancer. Treatment, tumor size and tumor status at second resection were independent variables associated with recurrence. Concomitant Cis was not predictive of failure of BCG therapy. Independent factor for treatment failure was remaining T1 stage at second resection. In Paper II &III we investigated the validity of emmprin, survivin and CCTα proteins as biomarkers for response and survival before neoadjuvant cisplatin chemotherapy. Bladder tumor specimens were obtained before therapy from a total of 250 patients with T1-T4 bladder cancer enrolled in 2 randomized trials comparing neoadjuvant chemotherapy before cystectomy with a surgery only arm. Protein expression was determined by immunohistochemistry (IHC). Patients in the chemotherapy cohort with negative emmprin and CCTα expression had significantly better overall survival (OS) than those with positive expression. In Paper IV primary end point was examining STMN1 as prognostic factor in bladder cancer.  Analysis was performed on three bladder cancer patient cohorts using IHC, western blot and a bladder cancer cell line. High levels of STMN1, expression correlated to shorter disease-specific survival and the growth and migration of the cells were significantly reduced when transfecting the cells with STMN1 siRNA. Conclusion Risk assessment and predictors of outcomes could help in individualized treatment and follow up.  Biomarkers will become more important for treatment choices in bladder cancer management.

Submitted by Filipe dos Santos (fsantos@pucsp.br) on 2016-09-16T18:28:01Z No. of bitstreams: 1 Denise Lemes Fernandes Neves.pdf: 2188363 bytes, checksum: 4a1bd7219349e1f8e991cf4c160a63d2 (MD5) / Made available in DSpace on 2016-09-16T18:28:01Z (GMT). No. of bitstreams: 1 Denise Lemes Fernandes Neves.pdf: 2188363 bytes, checksum: 4a1bd7219349e1f8e991cf4c160a63d2 (MD5) Previous issue date: 2016-04-04 / This master thesis aims to understand the user with intellectual diversity to establish the requirements needed for software development aimed to this significant portion of the world population. They are people with Syndromes, cerebral palsy, mental retardation, and other cognitive impairment. The part of this research that relates intellectual disability and software, there are no important findings on system requirements for the development of technology for the subject of study. A preliminary survey disclosed that technologies developed for "intellectual disability" tend to give emphasis on learning and teaching and, even when its creators argue that such technologies can develop the cognitive, help users memory and perception, they do not have clear and potential features that can in fact be attributed to disability. These are technologies that can be used by any user with difficulties in learning and not necessarily diagnosed with a cognitive disability and do not measure how much the cognitive of these people was changed after their use. To perform our study it will be used field research, diagnostic analysis, interviews with experts and research in specialized production. It will be pointed out important system requirements using theories of Human-Computer interface and Intelligent Systems, being a contribution for health, education and computing areas / Este trabalho busca conhecer o usuário com diversidade intelectual para estabelecer os requisitos necessários no desenvolvimento de software aplicado para essa significativa parcela da população mundial. Dela fazem parte pessoas com Síndromes, paralisia cerebral, retardo mental, entre outros comprometimentos cognitivos. No cenário da pesquisa que relaciona deficiência intelectual e software, não existem grandes apontamentos sobre requisitos do sistema para a criação de tecnologia para o sujeito de estudo. Um levantamento preliminar mostrou que as tecnologias desenvolvidas para a “deficiência intelectual” tendem a dar ênfase ao ensino-aprendizagem e, mesmo quando seus criadores defendem que tais tecnologias podem desenvolver o cognitivo, ajudar na memorização e percepção dos usuários, não apresentam características claras e potenciais que possam realmente ser atribuídas à deficiência. São tecnologias que podem ser utilizadas por qualquer usuário com dificuldades com a aprendizagem e não necessariamente com diagnóstico de deficiência cognitiva e também não mensuram o quanto o cognitivo destas pessoas foi alterado após seu uso. Para a realização de nosso estudo será aplicado pesquisa de campo, análise de diagnósticos, entrevistas com especialistas e pesquisas em material especializado. Serão pontuados importantes requisitos de sistemas usando teorias de interface Humano-Computador e Sistemas Inteligentes, sendo uma colaboração para as áreas da saúde, da educação e da computação

Submitted by PPG Ci?ncia da Computa??o (ppgcc@pucrs.br) on 2018-10-10T20:57:07Z No. of bitstreams: 1 EDUARDO GHIDINI_DIS.pdf: 8577965 bytes, checksum: 7ff48e36f96f7a9d7a545af88149f951 (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-10-17T11:12:35Z (GMT) No. of bitstreams: 1 EDUARDO GHIDINI_DIS.pdf: 8577965 bytes, checksum: 7ff48e36f96f7a9d7a545af88149f951 (MD5) / Made available in DSpace on 2018-10-17T11:21:19Z (GMT). No. of bitstreams: 1 EDUARDO GHIDINI_DIS.pdf: 8577965 bytes, checksum: 7ff48e36f96f7a9d7a545af88149f951 (MD5) Previous issue date: 2017-03-29 / The growth of data generated through the Internet increases the difficulty of gather relevant information for users. To minimize this, it is necessary to presented this information to the users in such a way that they can analyze it and extract useful information. In recent years, the use of narratives for the presentation of large volumes of data has been explored, considering its benefits. In this context, the present work aims to analyze and propose ways of presenting data, in narrative form, in the context of social networks, in order to support user interaction and data analysis. The work presents a model for the representation of narratives through visualizations, with data extracted from social networks, conceived from studies related to the areas of narratives and information visualization. The model was instantiated to enable its analysis through user studies. The studies indicated that the model has several applications, allowing the analysis of different types of data as well as its use by users with and without prior specific interests. We believe that the present work can support the construction of interactive narratives, through visualizations with data extracted from social networks, contemplating, not only the construction of the interface itself but, also, the mapping of the data to its visual representation. / O crescimento de dados gerados com o uso da Internet, faz com que, tamb?m, aumente a dificuldade de fazer com que informa??es relevantes cheguem aos usu?rios. Para que isso aconte?a, al?m do tratamento da informa??o, ? necess?rio que esta informa??o seja apresentada de tal forma que o usu?rio consiga analis?-la e extrair informa??es ?teis. Nos ?ltimos anos, vem se explorando o uso de narrativas para a apresenta??o de grandes volumes de dados, considerando-se os benef?cios desta forma de apresenta??o. Neste contexto, o presente trabalho objetiva analisar e propor formas de apresentar dados, em forma de narrativas, no contexto de redes sociais, a fim de apoiar a intera??o do usu?rio e sua an?lise dos dados. O trabalho apresenta um modelo para representa??o de narrativas por meio de visualiza??es, com dados extra?dos de redes sociais, concebido a partir de estudos relacionados ?s ?reas de narrativas e de visualiza??o de informa??es. O modelo foi instanciado para que viabilizasse a sua an?lise por meio de estudos com usu?rios. Os estudos indicam que o modelo tem variadas aplica??es, tanto no sentido de possibilitar a an?lise de diferentes tipos de dados, como, tamb?m, no que se refere ao uso por usu?rios com e sem interesse pr?vio em algum assunto espec?fico. Atrav?s do modelo proposto e do resultado de sua an?lise, acredita-se que o presente trabalho possa apoiar a constru??o de narrativas interativas neste contexto, contemplando, n?o somente a constru??o da interface em si, mas, tamb?m, o mapeamento dos dados para a sua representa??o visual.

The 12th International St Gallen conference on breast cancer (2011) proposed patient categorization for systemic therapy, based on intrinsic breast cancer subtypes, defined by imunohistochemistry (IHC) test results. Since this classification is based on semi-quantitative expression of IHC biomarker expression, an issue of defining and applying cutoff values remains. Essential improvement in the IHC testing has become possible with digital image analysis tools enabling quantitative evaluation of the IHC data. This study explores data obtained by digital image analysis methods applied to evaluate a comprehensive biomarker dataset (p53, AR, p16, BCL2, SATB1, HIF1) along with well established (ER, PR, HER2, Ki67) biomarkers. Also, an extensive set of genetic and epigenetic biomarkers has been tested. For the first time, the dataset of 10 IHC biomarkers, evaluated by digital analysis was explored by the means of factor analysis to establish the intrinsic factors of biological variation and informative value of IHC biomarkers and their combinatiions. The results also provided insights into the significance and combinatorial effects of the established and relatively new biomarkers (p16, SATB1, HIF1, Ki67/BCL2, etc.). / XII tarptautinėje St Gallen krūties vėžio konferencijoje (2011) Ekspertų komisijos priimta nauja pacientų klasifikacija sisteminei terapijai atlikti, paremta biologiniais krūties vėžio subtipais, kurie apibrėžiami imunohistocheminiu tyrimu (IHC). Tačiau ši nauja navikų klasifikacija iš esmės pagrįsta pusiau kiekybiniu biožymenų raiškos vertinimu, todėl išlieka aktuali ribinių verčių nustatymo problematika. Esminiai pokyčiai IHC tyrimų srityje galimi atsiradus skaitmeninio vaizdinimo technologijoms, leidžiančiomis IHC tyrimų rezultatus analizuoti kiekybiniais parametrais. Darbe naudojant skaitmeninį vaizdo analizės metodą atliktas išsamus biologinių žymenų tyrimas leido palyginti svarbių, tačiau nepakankamai ištirtų (p53, AR, p16, BCL2, SATB1, HIF1) IHC žymenų informatyvumą su esamų prognozinių žymenų (ER, PR, HER2, Ki67) rodikliais. Ištirtas platus genetinių ir epigenetinių krūties vėžio žymenų spektras. Pirmą kartą 10 IHC žymenų rinkinio, įvertinto skaitmeninės analizės būdu, duomenys panaudoti faktorinės analizės metodu nustatyti jų variacijų vidinius veiksnius, atskleidžiančius biologinius dėsningumus ir IHC žymenų bei jų derinių informatyvumą. Šios analizės rezultatai leido naujai įvertinti publikuotų krūties vėžio IHC žymenų bei jų derinių (p16, SATB1, HIF1, Ki67/BCL2 ir kt.) informatyvumą.

XII tarptautinėje St Gallen krūties vėžio konferencijoje (2011) Ekspertų komisijos priimta nauja pacientų klasifikacija sisteminei terapijai atlikti, paremta biologiniais krūties vėžio subtipais, kurie apibrėžiami imunohistocheminiu tyrimu (IHC). Tačiau ši nauja navikų klasifikacija iš esmės pagrįsta pusiau kiekybiniu biožymenų raiškos vertinimu, todėl išlieka aktuali ribinių verčių nustatymo problematika. Esminiai pokyčiai IHC tyrimų srityje galimi atsiradus skaitmeninio vaizdinimo technologijoms, leidžiančiomis IHC tyrimų rezultatus analizuoti kiekybiniais parametrais. Darbe naudojant skaitmeninį vaizdo analizės metodą atliktas išsamus biologinių žymenų tyrimas leido palyginti svarbių, tačiau nepakankamai ištirtų (p53, AR, p16, BCL2, SATB1, HIF1) IHC žymenų informatyvumą su esamų prognozinių žymenų (ER, PR, HER2, Ki67) rodikliais. Ištirtas platus genetinių ir epigenetinių krūties vėžio žymenų spektras. Pirmą kartą 10 IHC žymenų rinkinio, įvertinto skaitmeninės analizės būdu, duomenys panaudoti faktorinės analizės metodu nustatyti jų variacijų vidinius veiksnius, atskleidžiančius biologinius dėsningumus ir IHC žymenų bei jų derinių informatyvumą. Šios analizės rezultatai leido naujai įvertinti publikuotų krūties vėžio IHC žymenų bei jų derinių (p16, SATB1, HIF1, Ki67/BCL2 ir kt.) informatyvumą. / The 12th International St Gallen conference on breast cancer (2011) proposed patient categorization for systemic therapy, based on intrinsic breast cancer subtypes, defined by imunohistochemistry (IHC) test results. Since this classification is based on semi-quantitative expression of IHC biomarker expression, an issue of defining and applying cutoff values remains. Essential improvement in the IHC testing has become possible with digital image analysis tools enabling quantitative evaluation of the IHC data. This study explores data obtained by digital image analysis methods applied to evaluate a comprehensive biomarker dataset (p53, AR, p16, BCL2, SATB1, HIF1) along with well established (ER, PR, HER2, Ki67) biomarkers. Also, an extensive set of genetic and epigenetic biomarkers has been tested. For the first time, the dataset of 10 IHC biomarkers, evaluated by digital analysis was explored by the means of factor analysis to establish the intrinsic factors of biological variation and informative value of IHC biomarkers and their combinatiions. The results also provided insights into the significance and combinatorial effects of the established and relatively new biomarkers (p16, SATB1, HIF1, Ki67/BCL2, etc.).

In situ protein detection in human tissues using antibodies reveals the cellular protein localization, and affinity-based proteomic studies can help to discover proteins involved in the development of diseases. However, antibodies often suffer from cross-reactivity, and the lack of positive and negative tissue controls for uncharacterized proteins complicates the mapping of the proteome. The aim of this thesis is thus to improve the methodology for validating antibodies used for immunostaining on formalin-fixed paraffin-embedded tissues. Two of the papers include comparisons between mRNA-expression and immunostaining of corresponding protein. In paper I, ISH and IHC staining patterns were compared on consecutive TMA-slides. The study of well-characterized genes showed that ISH could be used for validation of antibodies. ISH was further used for antibody evaluation, and could validate four out of nine antibodies showing potentially interesting staining patterns. In paper III, transcriptomic data generated by RNA-sequencing were used to identify tissue specific expression in lymphohematopoietic tissues. An increased expression in one or more of these tissues compared to other tissue types was seen for 693 genes, and these were further compared to the staining patterns of corresponding proteins in tissues. Antibody labeling is necessary for many immunoassays. In paper II, two techniques for antibody-biotinylation were compared, aiming to find a stringent labeling method for antibodies used for immunostaining on TMAs. The ZBPA-method, binding specifically to Fc-part of antibodies, was found to be superior to the Lightning Link-biotinylation kit targeting amine groups, since labeling of amine groups on stabilizing proteins in the antibody buffer causes unspecific staining. The localization of the estrogen receptor beta (ERβ) in human normal and cancer tissues was studied in paper IV. Thorough evaluation of 13 antibodies using positive and negative control cell lines showed that only one antibody, PPZ0506, is specific for ERβ in all three immunoassays used. Contradictory to previously published data, tissue profiling using PPZ0506 showed that ERβ is expressed in a limited number of normal and cancer tissues. In conclusion, the present investigations present tools for validation of antibodies used for large-scale studies of protein expression in tissues.