Single-particle analysis (SPA) is a technique in which a protein or protein complex sample israpidly frozen (vitrified) to obtain a photograph (also called micrographs) of the structure atnear-atomic resolution by three-dimensional reconstruction.
It is mainly divided into sample screening by negative staining under normal temperature electron microscopy, preparation and optimization of biological samples, cryo sample preparation and screening of frozen samples, 300kV electron microscopy data collection, and three-dimensional reconstruction.
The schematic diagram of cryo-electron microscopy single particle analysis is as follows:
Suitable for three-dimensional reconstruction of soluble protein/membrane protein/macromolecule complex.
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