Cryo-electron tomography of thinned synapses

Description

Here we provide a protocol for cryo-electron tomography (cryo-ET) of thinned synapses within rat primary neuron cultures. This workflow relies on cryo-focused-ion-beam (FIB) milling to enable unrestricted access to synapses within neuronal cultures and to achieve samples sufficiently thin (~150 nm) lamellae for high resolution cryo-ET imaging. Synapse targeting with and without cryo-fluorescence light microscopy (cryo-FLM) correlation for FIB milling and cryo-ET is introduced.

Identifier (DOI)

10.17504/protocols.io.n92ldm188l5b/v1

Tags

Cryo-ET
In Vitro
Microscopy - electron
Rat

Team

Team Harper

Program

Collaborative Research Network

Theme

PD Functional Genomics

Meet the Authors

Anna Siegert, MSc

Key Personnel: Team Harper,

Universitätsmedizin Göttingen
Arsen Petrovic, PhD

Key Personnel: Team Harper,

Max Planck Institute of Biochemistry
Thanh Thao Do

External Collaborator
Florelle Domart

External Collaborator
Ruben Fernandez-Busnadiego, PhD

Co-PI (Core Leadership): Team Harper,

University of Gottingen

Cryo-electron tomography of thinned synapses

Output Details

Meet the Authors

Anna Siegert, MSc

Arsen Petrovic, PhD

Thanh Thao Do

Florelle Domart

Ruben Fernandez-Busnadiego, PhD

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