Quantitative measurements of lung microvessels would benefit characterization of vascular function and remodeling in pulmonary vascular diseases. Previous studies have evaluated the utility of micro-CT in conjunction with exogenous radiopaque silicone polymer injection (Microfil) to visualize vascular networks in whole organs. However, micro-CT resolutions are limited and Microfil perfusion may lead to incomplete vessel filling and vessel rupture. Optical coherence microscopy (OCM) enables depth-resolved volumetric imaging of tissue scattering with micron isotropic resolution and may be an alternative to micro-CT. Here, we present a novel method for quantitative measurements of lung vasculature using multi-volumetric OCM. Murine lungs were perfused with scattering contrast, fixed, and optically cleared. The lungs were then imaged using a custom-built OCM system with overlapping volumetric datasets and mosaicked in post-processing. OCM data was collected on a custom-built SD-OCT system and integrated with a control system to synchronize OCM data acquisition/archiving with three-axis motorized stages for multi-volumetric mosaicking. A Bessel illumination scheme was used to extend the Rayleigh range and depth-of-field by ~40% while maintaining high lateral resolution. A cleared lung lobe was imaged with 840 OCM volumes (7x12x10) that were acquired over an 8x13x1.43 mm slab with ~2 μm isotropic resolution. The resulting data was segmented in post-processing to quantify vessel diameters. We believe this proof-of-concept demonstrates the utility of our OCM and tissue preparation approach, which can be extended to compare microvasculature changes in entire lung lobes in animal models of pulmonary disease.
Kevin Z. Derby, Christa F Gaskill, Susan M. Majka, and Yuankai Kenny K. Tao, "Quantitative multi-volumetric imaging of lung vasculature using optical coherence microscopy (Conference Presentation)," Proc. SPIE 10867, Optical Coherence Tomography and Coherence Domain Optical Methods in Biomedicine XXIII, 108670Z (Presented at SPIE BiOS: February 05, 2019; Published: 4 March 2019); https://doi.org/10.1117/12.2511542.6008538139001.
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