Our research aims at quantifying pulmonary microvascular density in various mouse lung lobules considering anatomical differences and age-related changes using isolectin B4 staining and ImageJ analysis. Key questions that we are trying to address are how to quantify microvascular density in different lung lobules accurately, and what changes occur in microvascular density across lobules with aging. Recent advancements in pulmonary microvascular research, including improved tissue clearing and fixation techniques, advanced imaging, and software tools for accurate quantification.
These developments enhance our understanding of microvascular changes in aging and related diseases, like asthma and COVID-19, offering insights for potential therapies. The main challenges are accurately locating lesions, properly sectioning lung tissues, mitigating fixative-related tissue damage, and lack of specific markers for lung endothelial cells. Traditional methods often overlook the variations inside shape and vascularization across different lung lobes.
Our protocol addresses the gap by providing a comprehensive approach to sectioning lung lobes and quantifying microvascular density using isolectin before staining. The combination of IB4 staining with the free software ImageJ for advanced microvascular density analysis ensures accurate and useful results. The cellular and molecular mechanisms by which pulmonary microvascular lesions participate in the progression of lung-related diseases, and develop new strategies or targets for rescuing pulmonary microvascular function to promote lung repair and induce functional recovery.
