Micrometer-resolution X-ray tomographic full-volume reconstruction of an intact post-mortem juvenile rat lung

• Published in: Histochemistry and cell biology Vol. 155; no. 2; pp. 215 - 226
• Main Authors: Borisova, Elena, Lovric, Goran, Miettinen, Arttu, Fardin, Luca, Bayat, Sam, Larsson, Anders, Stampanoni, Marco, Schittny, Johannes C., Schlepütz, Christian M.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.02.2021; Springer Nature B.V
• Subjects: Animal models; Biochemistry; Biomedical and Life Sciences; Biomedicine; Cell Biology; Developmental Biology; Fast tomography; Gas exchange; Image reconstruction; Large volume tomography; Lung diseases; Lung imaging; Original Paper; Parenchyma; Spatial discrimination; Trachea; Visual field; X-ray tomography; X-rays; X-ray tomography; Large volume tomography; Fast tomography; Image reconstruction; Lung imaging
• ISSN: 0948-6143; 1432-119X; 1432-119X
• DOI: 10.1007/s00418-020-01868-8

In this article, we present an X-ray tomographic imaging method that is well suited for pulmonary disease studies in animal models to resolve the full pathway from gas intake to gas exchange. Current state-of-the-art synchrotron-based tomographic phase-contrast imaging methods allow for three-dimensional microscopic imaging data to be acquired non-destructively in scan times of the order of seconds with good soft tissue contrast. However, when studying multi-scale hierarchically structured objects, such as the mammalian lung, the overall sample size typically exceeds the field of view illuminated by the X-rays in a single scan and the necessity for achieving a high spatial resolution conflicts with the need to image the whole sample. Several image stitching and calibration techniques to achieve extended high-resolution fields of view have been reported, but those approaches tend to fail when imaging non-stable samples, thus precluding tomographic measurements of large biological samples, which are prone to degradation and motion during extended scan times. In this work, we demonstrate a full-volume three-dimensional reconstruction of an intact rat lung under immediate post-mortem conditions and at an isotropic voxel size of (2.75 µm) 3 . We present the methodology for collecting multiple local tomographies with 360° extended field of view scans followed by locally non-rigid volumetric stitching. Applied to the lung, it allows to resolve the entire pulmonary structure from the trachea down to the parenchyma in a single dataset. The complete dataset is available online ( https://doi.org/10.16907/7eb141d3-11f1-47a6-9d0e-76f8832ed1b2 ).