Single-molecule localization microscopy (SMLM) is a potent tool to examine biological systems with unprecedented resolution, enabling the investigation of increasingly smaller structures. At the forefront of these developments is DNA-based point accumulation for imaging in nanoscale topography (DNA-PAINT), which exploits the stochastic and transient binding of fluorescently labeled DNA probes. In its early stages the implementation of DNA-PAINT was burdened by low-throughput, excessive acquisition time, and difficult integration with live-cell imaging. However, recent advances are addressing these challenges and expanding the range of applications of DNA-PAINT. We review the current state of the art of DNA-PAINT in light of these advances and contemplate what further developments remain indispensable to realize live-cell imaging.

单分子定位显微镜 (SMLM) 是一种有效的工具，可以以前所未有的分辨率检查生物系统，从而能够研究越来越小的结构。这些发展的最前沿是基于 DNA 的纳米级形貌成像点积累 (DNA-PAINT)，它利用了荧光标记的 DNA 探针的随机和瞬时结合。在其早期阶段，DNA-PAINT 的实施受到低通量、过长的采集时间以及难以与活细胞成像集成的负担。然而，最近的进展正在解决这些挑战并扩大 DNA-PAINT 的应用范围。鉴于这些进展，我们回顾了 DNA-PAINT 的当前状态，并考虑了哪些进一步的发展对于实现活细胞成像仍然必不可少。