This paper describes how specimens of nervous tissue can be prepared for successful imaging in Xray Micro Computed Tomography (mu CT), 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 mu m x 100 mu m 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 mu CT 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. (C) 2010 Elsevier B.V. All rights reserved.
- X-ray micro computed tomography
- Rat sciatic nerve
- Osmium tetroxide
- Nerve scaffold
- Image analysis