Single fluorescent probes enabling simultaneous visualization of duple organelles: Design principles, mechanisms, and applications

[Display omitted] •A summary of single fluorescent probes for simultaneous visualization of duple organelles is devoted for the first rime.•This review describes the principles, strategies, advantages, possible shortages, and applications of these unique probes.•The future guidance and potential applications of these probes in cell biology and diagnosis are included. A unique kind of fluorescent probes which can target duple organelles have been continuously presented in very recent years. These probes have demonstrated great potential to visualize the interplay between different organelles and the visualization of invisible biophysical parameters. However, a systematic review on this special and significant topic is still absent, which may hinder the development of this area. In this review, we have systematically summarized these probes, and classified them in to three classes according to their biophysical applications. The first class involves probes to discriminate duple organelles in different emission colors, which showed potential applications to visualize the interplay between different organelles. The second class can change their subcellular localization upon the variation of biophysical parameters, which have demonstrated wide applications to visualize these invisible biophysical parameters. The third class may change their localization and concurrently shift their emission upon the variation of biophysical parameters, which enables simultaneous visualization of duple organelles and biophysical parameters. The mechanisms, design strategies, and biological applications of these probes were discussed, the prospect and possible future development of the area were finally included. We expected that this review can guide the development of probes targeting multiple organelles, and facilitate the studies on relative biomedical areas.