title: Large-volume reconstruction of brain tissue from high-resolution serial section images acquired by SEM-based scanning transmission electron microscopy.
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.
authors.affiliation: Center for Learning and Memory, The University of Texas at Austin, Austin, TX, USA.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Kuwajima
authors.forename: John M
authors.initials: JM
authors.lastname: Mendenhall
authors.forename: Kristen M
authors.initials: KM
authors.lastname: Harris
mesh_terms: D000818:Animals; D001921:Brain / Q000033:anatomy & histology* / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D017348:Microscopy, Electron, Scanning Transmission / Q000379:methods*; D008867:Microtomy; D009929:Organ Size; D010477:Perfusion; D051381:Rats; D013194:Staining and Labeling; D016707:Tissue Fixation