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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.

The mechanical properties of cells play an important role in cell development and function. Therefore, measurement of cell mechanical properties is a fundamental and essential tool for cell research. In this article, a novel method to estimate the force exerted on a living cell is proposed based on glass needle deformation, which does not require additional physical sensors compared with other force‐sensing methods. The three‐dimensional (3D) spatial state of the needle is reconstructed, and the parameters of needle deflection are obtained based on a multi‐focus image fusion algorithm. The average reconstruction error of this algorithm is 0.94 µm. Based on the deformation of the needle, a mechanical model of needle deformation is established, and the model is calibrated using a constructed calibration system. At the range of 0–200 µN, the highest resolution is 0.002 µN and the lowest resolution is 5.3 µN. The proposed method can be used to estimate the force exerted by a needle on the surface of a living cell.