Abstract
Optical projection tomography (OPT) is a powerful tool for three-dimensional imaging of mesoscopic biological samples with great use for biomedical phenotyping studies. We present a fluorescent OPT platform that enables direct visualization of biological specimens and processes at a centimeter scale with high spatial resolution, as well as fast data throughput and reconstruction. We demonstrate nearly isotropic sub-28 µm resolution over more than 60 mm3 after reconstruction of a single acquisition. Our setup is optimized for imaging the mouse gut at multiple wavelengths. Thanks to a new sample preparation protocol specifically developed for gut specimens, we can observe the spatial arrangement of the intestinal villi and the vasculature network of a 3-cm long healthy mouse gut. Besides the blood vessel network surrounding the gastrointestinal tract, we observe traces of vasculature at the villi ends close to the lumen. The combination of rapid acquisition and a large field of view with high spatial resolution in 3D mesoscopic imaging holds an invaluable potential for gastrointestinal pathology research.
Highlights
Mesoscopic imaging focuses on samples typically ranging from a few millimeters up to several centimeters in at least one dimension [1]
We described a new multispectral Optical projection tomography (OPT) setup optimized for the imaging of the mouse gut and capable of spanning 3 orders of magnitude in size along the 3 dimensions, from a few micrometers to centimeters
We showed that the adoption of a multispectral fluorescence approach on cleared samples subject to specific stainings allow to extract anatomic and structural information within a whole organ structure
Summary
Mesoscopic imaging focuses on samples typically ranging from a few millimeters up to several centimeters in at least one dimension [1]. Systematic longitudinal imaging of intact gut tissues is a desirable tool to advance our understanding of the fundamental interactions between the enteric nervous system, the immune system, the epithelial barrier and microbes Such imaging range can be exploited for the molecular diagnosis of Parkinson’s disease. Recent studies using conventional microscopy have revealed the presence of Lewy Bodies in the intestine, and highlighted their usefulness as early reliable biomarkers of the disease [2,3,4]. This method is challenging to apply in routine diagnostics because of poor tissue sampling in standard histological methods occurring when dealing with elongated specimens. The added advantages of using whole-tissue imaging are reliable and global biological marker characterization and quantification, decreasing the need for repeated endoscopic interventions
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