Size and specimen-dependent strategy for x-ray micro-ct and tem correlative analysis of nervous system samples

• Published in: Scientific reports Vol. 7; no. 1; pp. 2858 - 12
• Main Authors: Parlanti, P., Cappello, V., Brun, F., Tromba, G., Rigolio, R., Tonazzini, I., Cecchini, M., Piazza, V., Gemmi, M.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 06-06-2017; Nature Publishing Group; Nature Portfolio
• Subjects: 14/63; 59; 631/1647/245/1847; 631/1647/328/2082; 64/60; Animals; Biological samples; Computed tomography; Disease Models, Animal; Electron microscopy; Embedding; Humanities and Social Sciences; Imaging, Three-Dimensional; Leukodystrophy, Globoid Cell - diagnostic imaging; Leukodystrophy, Globoid Cell - pathology; Mice; Microscopy, Electron, Transmission; multidisciplinary; Nervous system; Nervous System - diagnostic imaging; Nervous System - pathology; Nervous System - ultrastructure; Rats; Science; Science (multidisciplinary); Spinal Cord - diagnostic imaging; Spinal Cord - pathology; Spinal Cord - ultrastructure; Transmission electron microscopy; X-Ray Microtomography - methods
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-017-02998-1

Correlative approaches are a powerful tool in the investigation of biological samples, but require specific preparation procedures to maintain the strength of the employed methods. Here we report the optimization of the embedding protocol of nervous system samples for a correlative synchrotron X-ray computed microtomography (micro-CT) and transmission electron microscopy (TEM) approach. We demonstrate that it is possible to locate, with the micrometric resolution of micro-CT, specific volumes of interest for a further ultrastructural characterization to be performed with TEM. This approach can be applied to samples of different size and morphology up to several cm. Our optimized method represents an invaluable tool for investigating those pathologies in which microscopic alterations are localized in few confined regions, rather than diffused in entire tissues, organs or systems. We present a proof of concept of our method in a mouse model of Globoid Cells Leukodistrophy.