Avaliação histomorfológica e imuno-histoquímica em amostras obtidas de autópsia para determinação de intervalo post mortem
Data
2022-05-26
Autores
Santos, Debora de Souza 
Orientadores
Iwamura, Edna Sadayo Miazato
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Objetivo. O objetivo deste estudo é analisar histomorfologicamente em Hematoxilina &
Eosina e avaliar marcadores imuno-histoquímicos em diferentes tecidos de um único
indivíduo, coletados em 6h, 60h e 80h post mortem, para estimativa de intervalo post
mortem. Metodologia. Neste estudo, Foram coletados 8 fragmentos amostrais, sendo
eles: músculo de parede abdominal (2), músculo cardíaco (2), fígado (2) e rim (2) sem
alterações patológicas visíveis macroscopicamente, obtidas em Janeiro de 2020, em
6h, 60h e 80h post mortem, divididos em grupos com soro e sem soro fisiológico. As
lâminas foram submetidas à imuno-histoquímica para a investigação de anticorpos
HepPar-1, RCC e HHF35, e realizou-se análise microscópica e elaboração de
histograma através do software ImageJ. Resultados. O processo autolítico está
presente em todas as lâminas, porém o avanço é visível ao longo do tempo. As
lâminas 6h foram consideradas como controle positivo, visto que a coleta e fixação
ocorreram no ato da autópsia e a preservação das estruturas está superior às demais
lâminas. A imuno-histoquimica teve coloração positiva em todas as amostras. O
histograma de cada imagem é condizente com a leitura do patologista, entretanto não
houve diferenças significativas entre o grupo com soro fisiológico e o grupo sem soro
fisiológico. No nosso contexto, os histogramas podem auxiliar a leitura de lâminas
microscópicas de tecidos homogêneos e marcadores citoplasmáticos na rotina da
patologia clínica. Conclusão. Lâminas coradas com Hematoxilina e Eosina são
essenciais para identificação tecidual. Tecidos homogêneos, como fígado e músculo,
são viáveis para estudos de intervalo post mortem e tecidos heterogêneos, como o
tecido renal, não possuem a mesma viabilidade devido à alta recorrência de autólise.
Consequentemente, os marcadores HepPar-1 e HHF35 são viáveis, porém, o
marcador RCC não apresenta boa viabilidade. Nossos resultados corroboram os
achados em literatura e podem fornecer informações relevantes para futuras
pesquisas.
Objective. The objective of this study is to evaluate immunohistochemical markers in different tissues from a single individual, collected up to 6h, 60h and 80h post mortem, to estimate the post mortem interval. Methodology. In this study, a histomorphological analysis was performed on different autopsy tissues to assess the pattern of protein degradation and potential immunohistochemical markers. In the present research, the identity of the autopsied participant was preserved, remaining anonymous at all stages of the study. Eight sample fragments were collected, namely: abdominal wall muscle (2), cardiac muscle (2), liver (2) and kidney (2) without macroscopically visible non-tumor and pathological changes, obtained in January 2020. The slides were submitted to immunohistochemistry for investigation of HepPar-1, RCC and HHF35 antibodies, submitted to microscopic and histogram analysis using ImageJ software. Results. The autolytic process is present in all blades, however the advance is visible over time. The T6 slides were considered as a positive control, since the collection and fixation took place during the autopsy and the preservation of the structures is superior to the other slides. The histogram of each image is consistent with the pathologist's reading, however, there were no significant differences between the saline group and the non-saline group. In our context, histograms can help the reading of microscopic slides of homogeneous tissues and cytoplasmic markers in the routine of clinical pathology. Conclusion. Slides stained with Hematoxylin and Eosin are essential for tissue identification, considering the decomposition rates that disfigure the tissue. Homogeneous tissues, such as liver and muscle, are viable for post mortem interval studies, and heterogeneous tissues, such as renal tissue, do not have the same viability due to the high recurrence of autolysis. Our results corroborate the findings in the literature and may provide relevant information for future research.
Objective. The objective of this study is to evaluate immunohistochemical markers in different tissues from a single individual, collected up to 6h, 60h and 80h post mortem, to estimate the post mortem interval. Methodology. In this study, a histomorphological analysis was performed on different autopsy tissues to assess the pattern of protein degradation and potential immunohistochemical markers. In the present research, the identity of the autopsied participant was preserved, remaining anonymous at all stages of the study. Eight sample fragments were collected, namely: abdominal wall muscle (2), cardiac muscle (2), liver (2) and kidney (2) without macroscopically visible non-tumor and pathological changes, obtained in January 2020. The slides were submitted to immunohistochemistry for investigation of HepPar-1, RCC and HHF35 antibodies, submitted to microscopic and histogram analysis using ImageJ software. Results. The autolytic process is present in all blades, however the advance is visible over time. The T6 slides were considered as a positive control, since the collection and fixation took place during the autopsy and the preservation of the structures is superior to the other slides. The histogram of each image is consistent with the pathologist's reading, however, there were no significant differences between the saline group and the non-saline group. In our context, histograms can help the reading of microscopic slides of homogeneous tissues and cytoplasmic markers in the routine of clinical pathology. Conclusion. Slides stained with Hematoxylin and Eosin are essential for tissue identification, considering the decomposition rates that disfigure the tissue. Homogeneous tissues, such as liver and muscle, are viable for post mortem interval studies, and heterogeneous tissues, such as renal tissue, do not have the same viability due to the high recurrence of autolysis. Our results corroborate the findings in the literature and may provide relevant information for future research.
Descrição
Citação
SANTOS, D.S. Avaliação histomorfológica e imuno-histoquímica em amostras obtidas de autópsia para determinação de intervalo Post Mortem. São Paulo, 2022. 76 f. Dissertação (Mestrado em Patologia) - Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP). São Paulo, 2022.