Nucleic Acid Probes for Single-Molecule Localization Imaging of Cellular Biomolecules

Endogenous biomolecules in cells are the basis of all life activities. Directly visualizing the structural characteristics and dynamic behaviors of cellular biomolecules is significant for understanding the molecular mechanisms in various biological processes. Single-molecule localization microscopy (SMLM) can circumvent the optical diffraction limit, achieving analysis of the fine structures and biological processes in living cells with nanoscale resolution. However, the large size of traditional imaging probes prevents SMLM from accurately locating fine structures and densely distributed biomolecules within cells. In recent years, nucleic acid probes have emerged as potential tools to replace conventional SMLM probes by virtue of their small size and high specificity. In addition, due to their programmability, nucleic acid probes with different conformations can be constructed via sequence design, further extending the application of SMLM in bioanalysis. Here, we discuss the design concepts of different conformational nucleic acid probes for SMLM and summarize the application of SMLM based on nucleic acid probes in the field of biomolecules. Furthermore, we provide a summary and future perspectives of the nucleic acid probe-based SMLM technology, aiming to provide guidance for the acquisition of nanoscale information about cellular biological processes.