Software-Assisted Image Analysis for Identification and Quantification of Sinusoidal Dilatation and Centrilobular Fibrosis in Masson’s Trichrome-Stained Liver Specimens
Douglas Mesadri Gewehr, Allan Fernando Giovanini, Seigo Nagashima, Ana Cristina Lira Sobral, Fernando Bermudez Kubrusly, Luiz Fernando Kubrusly.
Período de Execução:2020 – 2021
Área de Concentração:Hipertensão Arterial Pulmonar Experimental
Categoria:Iniciação Científica
Número Protocolo de Aprovação:CEUA/FEMPAR protocolo n. 2577/2020.
Recurso Financeiro:PIBIC/CNPq FEMPAR.
Resumo/Abstract:Digital image technologies have been introduced to pathology diagnosis, allowing an objective quantitative assessment, more accurate rather than instead of the semi-quantitative analysis. Few studies involving the quantitative measurement of software-assisted liver fibrosis have been published. Even scarcer are studies evaluating the centrilobular fibrosis and sinusoidal lumen area in liver Masson’s Trichrome-stained specimens. We propose a reliable, precise and computerized protocol to objectively quantify any amount of fibrosis and sinusoidal dilatation in liver Masson’s Trichrome-stained specimen. Once fibrosis had been established, liver samples were collected, histologically processed, stained with Masson’s trichrome, and whole-slide images were captured with an appropriated digital pathology slide scanner. After, a random selection of the regions of interest (ROI’s) was conducted. The data were subjected to software-assisted image analysis (ImageJ®). The analysis of 250 ROI’s allowed us to empirically obtain the best application settings to identify the centrilobular fibrosis (CF) and sinusoidal lumen (SL). After the establishment of the colour threshold application settings, an in-house Macro was recorded to set the measurements (fraction area and total area) and calculate the CF and SL ratios by an automatic batch processing. It provided an objective, reproducible and reliable method that precisely identifies and quantifies the area occupied by fibrous tissue and sinusoidal lumen in Masson’s trichrome-stained livers specimens. This technique could also be applied in the quantification of fibrosis of other liver zones and compartments. Therefore, to the best of our knowledge, no standardized sinusoidal lumen area quantification method has been described with Masson’s trichrome-stained livers specimens.
Keywords: liver fibrosis; sinusoidal segmentation; sinusoidal dilatation; image analysis; quantitative pathology; Masson’s trichrome-stained specimens.
Participação em Eventos:- II Congresso da Sociedade de Anatomia do Estado do Rio de Janeiro (SAERJ), 2021.
- Segundo Colocado Categoria Pôster II Congresso da Sociedade de Anatomia do Estado do Rio de Janeiro (SAERJ), 2021.