Block Face Scanning Electron Microscopy of Fluorescently Labeled Axons Without Using Near Infra-Red Branding.
Block Face Scanning Electron Microscopy of Fluorescently Labeled Axons Without Using Near Infra-Red Branding.
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DOI：
10.3389/fnana.2018.00088
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发表时间：
2018
影响因子：
2.9
通讯作者：
Knott G
中科院分区：
文献类型：
--
作者：
Maclachlan C;Sahlender DA;Hayashi S;Molnár Z;Knott G
在本文中，我们描述了一种方法，该方法允许荧光标记结构（如轴突）成为电子显微镜（EM）分析的目标，而无需将其标记转换为电子密集染色剂，引入任何基准标记或以高分辨率成像大体积。我们最佳地保存和染色脑组织进行超微结构分析，并使用自然地标，如细胞体和血管，来定位以前使用共聚焦显微镜成像的神经突。该方法依赖于固定后用光学显微镜拍摄的低和高倍率视图来捕获组织结构的信息，这些信息可用于确定序列EM图像中感兴趣的结构的位置。这里展示的例子是小鼠丘脑后内侧核中表达皮质丘脑轴突的td Tomato，用共聚焦显微镜在固定组织中成像，随后用连续块面EM （SBEM）进行可视化，并重建成3D模型进行分析。
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.
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影响因子：
2.5
作者：
MATHERS, LH
通讯作者：
MATHERS, LH
用于基于电子显微镜的连接组学的大批量En-Bloc染色。
影响因子：
16.6
作者：
Hua Y;Laserstein P;Helmstaedter M
通讯作者：
Helmstaedter M
影响因子：
64.8
作者：
Bock, Davi D.;Lee, Wei-Chung Allen;Kerlin, Aaron M.;Andermann, Mark L.;Hood, Greg;Wetzel, Arthur W.;Yurgenson, Sergey;Soucy, Edward R.;Kim, Hyon Suk;Reid, R. Clay
通讯作者：
Reid, R. Clay
影响因子：
2.9
作者：
Jorstad A;Blanc J;Knott G
通讯作者：
Knott G
影响因子：
3.7
作者：
Hoerder-Suabedissen, Anna;Korrell, Kim V.;Molnar, Zoltan
通讯作者：
Molnar, Zoltan