Abstract: Cryo-electron microscopy (cryo-EM) has become a widely used technology in biophysics. In recent years, with the application of the direct electron detector device camera and continuous innovations in the single-particle reconstruction algorithms, cryo-EM has made tremendous progress. Macromolecular structures can now be determined with near atomic resolution (<4 Å) with this technique. As more and more proteins and complexes with important biomedical implications are being revealed with high-resolution, structural biology is undergoing a gradual revolution. Cryo-EM is attracting worldwide attention. In this article we describe its development and single-particle analysis techniques, taking the inflammasome complex and splicesome as examples to demonstrate the latest advances.