Second harmonic generation and multiphoton excited fluorescence microscopy methods were used to examine structural remodeling of the extracellular matrix in human lung alveolar walls undergoing emphysematous destruction. Fresh lung samples removed from a patient undergoing lung transplantation for very severe chronic obstructive pulmonary disease were compared to similar samples from an unused donor lung that served as a control. The generated spatially resolved 3D images show the spatial distribution of collagen, elastin and other endogenously fluorescent tissue components such as macrophages. In the case of control lung tissue, we found well ordered alveolar walls with composite type structure made up of collagen matrix and relatively fine elastic fibers. In contrast, lung tissue undergoing emphysematous destruction was highly disorganized with increased alveolar wall thickness compared to control lung tissue.