Automated Computer-Assisted Image Analysis for the Fast Quantification of Kidney Fibrosis

SIMPLE SUMMARY: Chronic kidney disease is a health problem in which the kidneys cannot function normally. Thus, they cannot filter blood effectively and cause waste accumulation in the organism, leading to serious health problems. Researchers use animals as models to replicate the human body’s behavior to understand this disease. In these studies, it is essential to evaluate the percentage of fibrosis (growth of fibrotic tissue similar to a scar in response to damage) to know the degree of kidney damage. Some researchers use programs to make the evaluation of fibrosis easier. However, this analysis is time-consuming because it needs to be made one image at a time and there are hundreds of samples in an animal model study. Here, we explain a method to conduct the same analysis but in a faster automated way with the assistance of a computer and a software package called CellProfiler™. The percentage of fibrosis using CellProfiler™ is similar to that obtained with the most widely used software for this kind of analysis called ImageJ. With the help of this approach, researchers can make more studies faster and easier and find new antifibrogenic therapies to address the common and worldwide health problem caused by chronic kidney disease. ABSTRACT: Chronic kidney disease (CKD) is a common and worldwide health problem and one of the most important causes of morbidity and mortality. Most primary research on this disease requires evaluating the fibrosis index in animal model kidneys, specifically using Masson’s trichrome stain. Different programs are used to calculate the percentage of fibrosis; however, the analysis is time-consuming since one image must be performed at a time. CellProfiler™ is a program designed to analyze data obtained from biological samples and can process multiple images through pipelines, and the results can be exported to databases. This article explains how CellProfiler™ can be used to automatically analyze kidney histology photomicrographs from samples stained with Masson’s trichrome stain to assess the percentage of fibrosis in an experimental animal model of CKD. A pipeline was created to analyze Masson’s trichrome-stained slides in a model of CDK induced by adenine at doses of 50 mg/kg and 100 mg/kg, in addition to samples with the vehicle (75% glycerin). The results were compared with those obtained by ImageJ, and no significant differences were found between both programs. The CellProfiler™ pipeline made here is a reliable, fast, and easy alternative for kidney fibrosis analysis and quantification in experimental animal models.