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This paper describes how specimens of nervous tissue can be prepared for successful imaging in X-ray Micro Computed Tomography (microCT), and how this method can be used to study the integration of nervous tissue into a polymeric scaffold. The sample preparation involves staining the biological tissue with osmium tetroxide to increase its X-ray attenuation, and a technique for maintaining the specimen in a moist environment during the experiment to prevent drying and shrinkage. Using this method it was possible to observe individual nerve fascicles and their relationship to the 3-D tissue structure. A scaffold supporting a regenerated sciatic nerve was similarly stained to distinguish the nervous tissue from the scaffold, and to observe how the nerve grew through a 2.5 mm long, 100 microm x 100 microm cross-section channel polyimide array. Furthermore, blood vessels could be identified in these images, and it was possible to monitor how a large proximal blood vessel split through the channel scaffold and proceeded down individual channels. This paper explains how microCT is a useful tool both for studying the location and extent of growth into a polymeric scaffold, and for determining whether the regenerated tissue has blood supply.

Original publication

DOI

10.1016/j.jneumeth.2010.01.032

Type

Journal

J Neurosci Methods

Publication Date

30/04/2010

Volume

188

Pages

39 - 44

Keywords

Animals, Biocompatible Materials, Guided Tissue Regeneration, Image Processing, Computer-Assisted, Nerve Regeneration, Rats, Sciatic Nerve, Tissue Engineering, Tissue Scaffolds, X-Ray Microtomography