Developing and utilizing the crystal ribcage platform to probe respiratory and circulatory functions of the lung from whole organ to single alveolus in health and disease
Embargo Date
2027-05-28
OA Version
Citation
Abstract
Understanding pulmonary disease pathogenesis and therapeutic response requires probing the lung at cellular resolution during active respiration and circulation. However, real-time optical imaging of the intact lung has remained challenging due to the dynamic motion of the lung during physiological breathing and the delicate air-liquid interface. To address these challenges, we developed the “crystal ribcage”, a transparent ribcage that enables multiscale optical imaging of the functioning lung from the whole-organ to single-cell level. The crystal ribcage preserves the three-dimensional architecture, air–liquid interface, cellular diversity, and respiratory–circulatory functions of the lung while enabling controlled modulation of lung biophysics and immunity through intravascular, intrapulmonary, intraparenchymal, and optogenetic interventions. Using this platform in murine models of pulmonary disease, we directly visualized remodeling of respiratory and vascular transport processes at the level of individual alveoli and capillaries during disease progression. We further applied the crystal ribcage to investigate the transport of inhaled aerosols carrying pathogens, pollutants, therapeutics, and diagnostic agents within functional alveoli during active ventilation. Real-time imaging of single aerosol droplets revealed deterministic heterogeneity in aerosol deposition at both intra- and inter-alveolar scales. Aerosols formed a characteristic “mosaic pattern”, in which only specific clusters of alveoli received particles. This pattern emerged consistently during spontaneous breathing and under controlled ventilation, across diverse particle types including small molecules, nanobodies, nanoparticles, microplastics, therapeutics, and pathogens. Mosaic deposition was also observed in murine, porcine, and human lungs and evolved with aging in mice. Together, these findings establish the crystal ribcage as a powerful platform for intravital lung imaging and reveal previously unrecognized spatial heterogeneity in aerosol transport with important implications for respiratory health and disease. Building on the crystal ribcage, we also developed an intravital lung imaging window that preserves both breathing and perfusion function in vivo, enabling longitudinal studies of pulmonary function, disease progression, and response to therapies over multiple days. This platform preserves spontaneous breathing and allows repeated imaging of the same alveoli and capillaries over time, providing a unique tool to track disease progression, immune cell dynamics, and aerosol transport under physiologically relevant conditions.
Description
2026
License
Attribution-NoDerivatives 4.0 International