3D Analysis of Synaptic Ultrastructure in Organotypic Hippocampal Slice Culture by High-Pressure Freezing and Electron Tomography

Cordelia Imig; Benjamin H Cooper

doi:10.1007/978-1-4939-6688-2_15

3D Analysis of Synaptic Ultrastructure in Organotypic Hippocampal Slice Culture by High-Pressure Freezing and Electron Tomography

Cordelia Imig, Benjamin H Cooper

Research output: Contribution to journal › Journal article › Research

7 Citations (Scopus)

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.

Original language	English
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	1538
Pages (from-to)	215-231
Number of pages	17
ISSN	1064-3745
DOIs	https://doi.org/10.1007/978-1-4939-6688-2_15
Publication status	Published - 2017
Externally published	Yes

Keywords

Animals
Electron Microscope Tomography/methods
Hippocampus/cytology
Image Processing, Computer-Assisted
Imaging, Three-Dimensional/methods
Mice
Microscopy, Electron, Transmission/methods
Synapses/ultrastructure

Access to Document

10.1007/978-1-4939-6688-2_15

Cite this

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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.",

keywords = "Animals, Electron Microscope Tomography/methods, Hippocampus/cytology, Image Processing, Computer-Assisted, Imaging, Three-Dimensional/methods, Mice, Microscopy, Electron, Transmission/methods, Synapses/ultrastructure",

author = "Cordelia Imig and Cooper, {Benjamin H}",

year = "2017",

doi = "10.1007/978-1-4939-6688-2_15",

language = "English",

volume = "1538",

pages = "215--231",

journal = "Methods in Molecular Biology",

issn = "1064-3745",

publisher = "Humana Press",

}

TY - JOUR

T1 - 3D Analysis of Synaptic Ultrastructure in Organotypic Hippocampal Slice Culture by High-Pressure Freezing and Electron Tomography

AU - Imig, Cordelia

AU - Cooper, Benjamin H

PY - 2017

Y1 - 2017

N2 - 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.

AB - 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.

KW - Animals

KW - Electron Microscope Tomography/methods

KW - Hippocampus/cytology

KW - Image Processing, Computer-Assisted

KW - Imaging, Three-Dimensional/methods

KW - Mice

KW - Microscopy, Electron, Transmission/methods

KW - Synapses/ultrastructure

U2 - 10.1007/978-1-4939-6688-2_15

DO - 10.1007/978-1-4939-6688-2_15

M3 - Journal article

C2 - 27943193

VL - 1538

SP - 215

EP - 231

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

SN - 1064-3745

ER -