Single-cell patterning technology has revealed significant contributions of single cells to conduct basic and applied biological studies in vitro such as the understanding of basic cell functions, neuronal network formation, and drug screening. Unlike traditional population-based cell patterning approaches, single-cell patterning is an effective technology of fully understanding cell heterogeneity by precisely controlling the positions of individual cells. Therefore, much attention is currently being paid to this technology, leading to the development of various micro-nanofabrication methodologies that have been applied to locate cells at the single-cell level. In recent years, various methods have been continuously improved and innovated on the basis of existing ones, overcoming the deficiencies and promoting the progress in biomedicine. In particular, microfluidics with the advantages of high throughput, small sample volume, and the ability to combine with other technologies has a wide range of applications in single-cell analysis. Here, we present an overview of the recent advances in single-cell patterning technology, with a special focus on current physical and physicochemical methods including stencil patterning, trap- and droplet-based microfluidics, and chemical modification on surfaces via photolithography, microcontact printing, and scanning probe lithography. Meanwhile, the methods applied to biological studies and the development trends of single-cell patterning technology in biological applications are also described.
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