Numerous pulmonary disorders are associated with fibrosis, however, current methodology falls short when studying turbid tissue. Optical clearing (OC) offers a chemical-based approach for deep tissue imaging. We optimized organ-level OC approach and combined it with large-scale, label-free multiphoton microscopy (MPM) and second harmonic generation microscopy (SHGM) to reveal fibrillar collagen in whole murine lungs. The standardization of several underlying steps allowed for a streamline approach. This method revealed significant differences in collagen deposition between control and treated lungs, while establishing a new approach based on OC and MPM/SHGM imaging for 3D analysis of lung fibrosis in the whole lung.
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