Imaging peripheral nerve micro-anatomy with MUSE, 2D and 3D approaches

Author:

Kolluru Chaitanya,Todd Austin,Upadhye Aniruddha R.,Liu Yehe,Berezin Mikhail Y.,Fereidouni Farzad,Levenson Richard M.,Wang Yanming,Shoffstall Andrew J.,Jenkins Michael W.,Wilson David L.

Abstract

AbstractUnderstanding peripheral nerve micro-anatomy can assist in the development of safe and effective neuromodulation devices. However, current approaches for imaging nerve morphology at the fiber level are either cumbersome, require substantial instrumentation, have a limited volume of view, or are limited in resolution/contrast. We present alternative methods based on MUSE (Microscopy with Ultraviolet Surface Excitation) imaging to investigate peripheral nerve morphology, both in 2D and 3D. For 2D imaging, fixed samples are imaged on a conventional MUSE system either label free (via auto-fluorescence) or after staining with fluorescent dyes. This method provides a simple and rapid technique to visualize myelinated nerve fibers at specific locations along the length of the nerve and perform measurements of fiber morphology (e.g., axon diameter and g-ratio). For 3D imaging, a whole-mount staining and MUSE block-face imaging method is developed that can be used to characterize peripheral nerve micro-anatomy and improve the accuracy of computational models in neuromodulation. Images of rat sciatic and human cadaver tibial nerves are presented, illustrating the applicability of the method in different preclinical models.

Funder

National Institute of Biomedical Imaging and Bioengineering

National Institutes of Health

National Cancer Institute

Publisher

Springer Science and Business Media LLC

Subject

Multidisciplinary

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