Software-Assisted Image Analysis for Identification and Quantification of Hepatic Sinusoidal Dilatation and Centrilobular Fibrosis

Abstract

Background: Heart dysfunction and liver disease often coexist because of systemic disorders. Any cause of right ventricular failure may precipitate hepatic congestion and fibrosis. Digital image technologies have been introduced to pathology diagnosis, allowing an objective quantitative assessment. The quantification of fibrous tissue in liver biopsy sections is extremely important in the classification, diagnosis and grading of chronic liver disease. Aim: To create a semi-automatic computerized protocol to quantify any amount of centrilobular fibrosis and sinusoidal dilatation in liver Masson’s Trichrome-stained specimen. Method: 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®). Results: The analysis of 250 ROI’s allowed 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. Conclusion: Was possible to create a more detailed method that identifies and quantifies the area occupied by fibrous tissue and sinusoidal lumen in Masson’s trichrome-stained livers specimens.

Hypertension secondary to nitric oxide depletion produces oxidative imbalance and inflammatory/fibrotic outcomes in the cornea of C57BL/6 mice

“…Furthermore, collagen fibers in the corneas were visualized using Masson’s trichrome (MTS; Merk, Germany, 1.00485) and picro Sirius Red (SRS; Sigma-Aldrich Inc, St Louis, USA) staining. MTS and SRS pictures were quantified using Image J-NIH freeware (v. 2.0.0) and following protocols detailed elsewhere [ 17 ].…”

Automated whole slide image analysis for a translational quantification of liver fibrosis

“…Digital pathology has considerably developed over the past years with the following advantages: to provide a continuous measurement of histological endpoints, independency from the reader’s subjectivity. Computerized image techniques were already applied to histological liver section in various fibrotic rodent models 13 – 15 and to human biopsies 16 – 18 . One of the first applications of digital pathology in the field of hepatology was liver fibrosis assessment.…”

Phosphotungstic Acid–treated Picrosirius Red Staining Improves Whole-slide Quantitative Analysis of Collagen in Histological Specimens

“…7 In addition, Sirius Red staining is a selective dye for collagen fibers 8 and has been established as the most stable method of visualizing fibrosis in liver biopsies. 9 Computer-assisted image analysis (CAIA), as well as Masson's trichrome, [10][11][12] reticulin, 13 Sirius Red, [14][15][16] and Elastica van Gieson staining, [17][18][19] is used widely for liver fibrosis assessment.…”