Abstract
The study of biological mesoscopic samples (those with characteristic dimensions of 1-10 mm) benefits from techniques that can analyse and depict anatomical information on a whole organism level. Optical Projection Tomography (OPT) uses visible light to generate 3D imaging data of transparent mesoscopic objects (e.g. cleared mouse embryos) at micron-level resolution in both fluorescence and transmission. We present a low-cost OPT solution to image large transparent specimens (up to 17 x 14 mm) at a lateral resolution of ∼23 um and provide an extensive open-source library for image calibration, reconstruction, and noise-reduction. In the development of treatment which induces alveolar cell proliferation and repair in the porcine pancreatic elastase (PPE) mouse model of emphysema, the use of OPT enables quantitative imaging of relevant proliferative cell lineage in cleared, large murine lung samples. The volume and distribution of alveolar cell clusters which have been labelled by immunofluorescence are measured in the intact lung of a mouse and visualized in 3D. Our design is easy to implement and is completely open-source, providing opportunities for further open microscopy developments for large organ imaging in model organisms.
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