title: 3D Analysis of Synaptic Ultrastructure in Organotypic Hippocampal Slice Culture by High-Pressure Freezing and Electron Tomography.
abstract: Transmission electron microscopy serves as a valuable tool for synaptic structure-function analyses aimed at identifying morphological features or modifications associated with specific developmental stages or dysfunctional synaptic states. By utilizing cryo-preparation techniques to minimize the introduction of structural artifacts during sample preparation, and electron tomography to reconstruct the 3D ultrastructural architecture of a synapse, the spatial organization and morphological properties of synaptic organelles and subcompartments can be quantified with unparalleled precision. In this chapter, we present an experimental approach combining organotypic slice culture, high-pressure freezing, automated freeze-substitution, and electron tomography to investigate spatial relationships between synaptic vesicles and active zone release sites in synapses from lethal mouse mutants.
authors.affiliation: Department of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, 37075, Göttingen, Germany.
authors.forename: Cordelia
authors.initials: C
authors.lastname: Imig
authors.affiliation: Department of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, 37075, Göttingen, Germany. cooper@em.mpg.de.
authors.forename: Benjamin H
authors.initials: BH
authors.lastname: Cooper
mesh_terms: D000818:Animals; D055032:Electron Microscope Tomography / Q000379:methods*; D006624:Hippocampus / Q000166:cytology* / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D046529:Microscopy, Electron, Transmission / Q000379:methods; D013569:Synapses / Q000648:ultrastructure*
keywords: Automated freeze-substitution (AFS); Electron tomography (ET); High-pressure freezing (HPF); Organotypic slice culture; Synaptic ultrastructure; Transmission electron microscopy (TEM)