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Spectral Reflectometric Microscopy on Myelinated Axons In Situ
Published on: July 2, 2018
Spectral Reflectometric Microscopy on Myelinated Axons In Situ
Junhwan Kwon1, Myunghwan Choi2
1Department of Biomedical Engineering, Sungkyunkwan University; Center for Neuroscience Imaging Research, Institute for Basic Science (IBS).
Abstract:
In a mammalian nervous system, myelin provides an electrical insulation by enwrapping the axon fibers in a multilayered spiral. Inspired by its highly-organized subcellular architecture, we recently developed a new imaging modality, named spectral reflectometry (SpeRe), which enables unprecedented label-free nanoscale imaging of the live myelinated axons in situ. The underlying principle is to obtain nanostructural information by analyzing the reflectance spectrum of the multilayered subcellular structure. In this article, we describe a detailed step-by-step protocol for performing a basic SpeRe imaging of the nervous tissues using a commercial confocal microscopic system, equipped with a white-light laser and a tunable filter. We cover the procedures of sample preparation, acquisition of spectral data, and image processing for obtaining nanostructural information.
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