Nanoimaging pp 253-273 | Cite as

Large-Volume Reconstruction of Brain Tissue from High-Resolution Serial Section Images Acquired by SEM-Based Scanning Transmission Electron Microscopy

  • Masaaki Kuwajima
  • John M. Mendenhall
  • Kristen M. Harris
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 950)

Abstract

With recent improvements in instrumentation and computational tools, serial section electron microscopy has become increasingly straightforward. A new method for imaging ultrathin serial sections is developed based on a field emission scanning electron microscope fitted with a transmitted electron detector. This method is capable of automatically acquiring high-resolution serial images with a large field size and very little optical and physical distortions. In this chapter, we describe the procedures leading to the generation and analyses of a large-volume stack of high-resolution images (64 μm × 64 μm × 10 μm, or larger, at 2 nm pixel size), including how to obtain large-area serial sections of uniform thickness from well-preserved brain tissue that is rapidly perfusion-fixed with mixed aldehydes, processed with a microwave-enhanced method, and embedded into epoxy resin.

Key words

3D reconstruction Hippocampus Microwave-enhanced processing Neuropil Perfusion Scanning electron microscopy Scanning transmission electron microscopy Serial sections Structural plasticity Ultramicrotomy 

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Masaaki Kuwajima
    • 1
  • John M. Mendenhall
    • 1
  • Kristen M. Harris
    • 1
    • 2
  1. 1.Center for Learning and MemoryThe University of Texas at AustinAustinUSA
  2. 2.Section of NeurobiologyThe University of Texas at AustinAustinUSA

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