Diamond Light Source, Harwell Science and Innovation Campus, Fermi Ave, Didcot OX11 0DE, UK.
Mary Lyon Centre at MRC Harwell, Harwell Campus, Becquerel Ave, Didcot OX11 ORD, UK.
Int J Mol Sci. 2023 Oct 14;24(20):15191. doi: 10.3390/ijms242015191.
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.
近年来，随着冷冻电子断层扫描（cryo-ET）的日益普及，人们一直在努力建立一种在电子显微镜网格（EM 网格）上直接培养原代神经元的方法。在这里，我们描述了一种在 EM 网格上建立成熟神经元网络的简单方法，其中包括形成突触接触。这些突触很薄，足以在不进行预先聚焦离子束铣削的情况下，在 Titan Krios 上直接观察到将突触小泡（SV）固定到活性区质膜的小细丝，如 SNARE 蛋白。
