An efficient protocol of cryo-correlative light and electron microscopy for the study of neuronal synapses

• 来源期刊: 生物物理学报
• 年，卷（期）: 2019,（3）
• 页码范围: 111-122
• 页数: 12页
• 语种: 中文

摘要

Cryo-electron tomography is an emerging electron microscopy technique for determining three-dimensional structures of cellular architectures near their native state at nanometer resolution,with a shortcoming of lack of specific labels.Fluorescence light microscopy,on the other hand,specifically visualizes target cellular and molecular components with fluorescent labels,but is limited to a resolution of tens to hundreds nanometers.Combining the advantages of the two techniques,we have developed a cryo-correlative light and electron microscopy system.Our system consists a custom-designed cryo-chamber that allows for fluorescence imaging of frozen-hydrated samples,and an algorithm to achieve accurate correlation.With this system and our optimized protocol,high-quality tomograms of neuronal synapses labelled by specific fluorescent tags in cultured hippocampal neurons are obtained at high efficiency.