Organic fluorescent probes for stochastic optical reconstruction microscopy (STORM): Recent highlights and future possibilities

Super-resolution fluorescence imaging by single-molecule localization microscopy (SMLM) offers the possibility of microscopic images with sub-diffraction spatial resolution. Stochastic optical reconstruction microscopy (STORM) is one of the emerging SMLM techniques that has contributed new insights into both the structures and functions of sub-cellular organelles in the cellular context with a spatial resolution virtually at the molecular level. Photo-switching of single fluorophores and position determination are the most common features of this SMLM technique, which allows molecule-resolved information as well as super-resolved images. However, achieving successful STORM-based images relies on the suitable choice of a fluorophore. In particular, the use of ideal organic fluorescent probes has great potential to circumvent common difficulties that arise during the construction of STORM images. However, there is hardly any comprehensive review that critically assesses the criteria for choosing ideal fluorescent probes for STORM and designing new efficient organic fluorescent probes to date. Therefore, this review has particularly focused on the choice of organic fluorescent probes, the essential features for designing new probes and the future prospects for resolving persistent issues in STORM imaging. The utility of organic fluorescent probes in multicolor STORM, 3D STORM and live cell STORM imaging are also discussed to provide a perspective concerning the true application potential of commonly used fluorescent dyes. In this review, we not only describe how organic fluorescent dyes have contributed to the growth of STORM-based super-resolution imaging in eukaryotic biology, but we also attempt to provide a basis on which advanced organic fluorescent probes can be designed and developed in the near future.