Single‐cell manipulation technology is an essential method of cell research, with wide applications in biological engineering, medical engineering, agricultural engineering, and other precise manipulation fields. Cell pose adjustment and cell puncture are considered as two basic operations of single‐cell manipulation. Cell pose adjustment can be used for cell sorting, cell trapping, cell orientation, cell transportation, etc. It consists of direct contact manipulation methods and noncontact manipulation methods (e.g., mechanical contact method, electric field, optical field, magnetic field, acoustic field, and hydrodynamic methods). Cell puncture consists of ordinary glass needle puncture methods, piezoelectric‐assisted puncture methods, and rotational oscillatory drill puncture methods. It aims to achieve directional and quantified injection or sampling with minimal damage. Herein, the recent research progress on single‐cell pose adjustment and puncture methods is systematically summarized and discussed, which involves the basic mechanism, characteristics, limitations, and applications. Some potential directions for future research are proposed in accordance with the above analysis. It is hoped that this review can provide a reference for academic research and industrial applications of single‐cell manipulation technology.