As one of the fastest growing frontier discipline, cryo-Electron Microscope （cryo-EM） has been emerging as the most powerful tool in determining the high resolution structures of bio-macromolecules. Our aim is to teach the basic principles and methods in cryo-Electron Microscope, especially Single Particle Analysis. With this course, we will present the basic knowledge of the Single Particle Analysis, the principle of negative staining Electron Microscope and cryo-EM sample preparation, to teach students to apply the knowledges and skills to solve biomolecular structures and biomedical problems.