Shuichi Hayashi    Department   Kawasaki Medical School  Kawasaki Medical School, Department of Anatomy,    Position   Associate Professor
Article types	原著
Language	English
Peer review	Peer reviewed
Title	Block face scanning electron microscopy of fluorescently labelled axons without using near infra-red branding.
Journal	Formal name：Frontiers in Neuroanatomy
Domestic / Foregin	Foregin
Volume, Issue, Page	12,pp.88
Author and coauthor	Maclachlan, C., Sahlender, D., Hayashi, S., Molnar, Z., Blanc, J. and Knott, G.
Publication date	2018
Summary	In this article, we describe the method that allows fluorescently tagged structures such as axons to be targeted for electron microscopy (EM) analysis without the need to convert their labels into electron dense stains, introduce any fiducial marks, or image large volumes at high resolution. We optimally preserve and stain the brain tissue for ultrastructural analysis and use natural landmarks, such as cell bodies and blood vessels, to locate neurites that had been imaged previously using confocal microscopy. The method relies on low and high magnification views taken with the light microscope, after fixation, to capture information of the tissue structure that can later be used to pinpoint the position of structures of interest in serial EM images. The examples shown here are td Tomato expressing cortico-thalamic axons in the posteromedial nucleus of the mouse thalamus, imaged in fixed tissue with confocal microscopy, and subsequently visualized with serial block-face EM (SBEM) and reconstructed into 3D models for analysis.