The airway mucosa is the primary tissue site exposed to inhaled particulate matter, which includes pathogens and allergens. While most inhaled particles are eliminated from the airways via mucociliary clearance, some pathogens may penetrate the mucosal epithelial barrier and an effective activation of the mucosal immune system is required to prevent further pathogen spread. Similarly, inhaled environmental allergens may induce an aberrant activation of immune cells in the airway mucosa, causing allergic airway disease. During the last years, several investigators employed advanced microscopic imaging on both intravital and tissue explant preparations to observe the dynamic behavior of various immune cells within their complex tissue environment. In the respiratory tract, most imaging studies focused on immune responses of the alveolar compartment in the lung periphery.

However, equally important immunological events occur more proximally in the mucosa of the conducting airways, both during infection and allergic responses, calling for a more detailed imaging analysis also at this site. In this review, I will outline the technical challenges of designing microscopic imaging experiments in the conducting airways and summarize our recent efforts in understanding airway mucosal immune cell dynamics in steady–state conditions, during infection and allergy.