Completing the canvas: advances and challenges for DNA-PAINT super-resolution imaging

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 DNAbased 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 livecell 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.

Automated summary

Single-molecule localization microscopy (SMLM) is a powerful tool for examining biological systems with unprecedented resolution. DNA-based point accumulation for imaging in nanoscale topography (DNA-PAINT) has faced challenges such as low throughput, long acquisition time, and difficulty integrating with live-cell imaging, but recent advances are addressing these issues and expanding its applications. The current state of DNA-PAINT is reviewed in light of these advancements, with consideration for further developments needed to achieve live-cell imaging.