Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 24, Issue 20, Pages -Publisher
MDPI
DOI: 10.3390/ijms242015191
Keywords
primary neurons; astrocytes; cryo-electron microscopy; synapse
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This article describes a straightforward method to establish a mature neuronal network on electron microscopy grids and visualize the formation of synaptic contacts.
With the increasing popularity of cryo-electron tomography (cryo-ET) in recent years, the quest to establish a method for growing primary neurons directly on electron microscopy grids (EM grids) has been ongoing. Here we describe a straightforward way to establish a mature neuronal network on EM grids, which includes formation of synaptic contacts. These synapses were thin enough to allow for direct visualization of small filaments such as SNARE proteins tethering the synaptic vesicle (SV) to the active zone plasma membrane on a Titan Krios without prior focused ion-beam milling.
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