ハヤシ シュウイチ
  林 周一
   所属   川崎医科大学  医学部 基礎医学 解剖学
   職種   准教授
論文種別 原著
言語種別 英語
査読の有無 査読あり
表題 Block face scanning electron microscopy of fluorescently labelled axons without using near infra-red branding.
掲載誌名 正式名:Frontiers in Neuroanatomy
掲載区分国外
巻・号・頁 12,88頁
著者・共著者 Maclachlan, C., Sahlender, D., Hayashi, S., Molnar, Z., Blanc, J. and Knott, G.
発行年月 2018
概要 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.