Targeted Conversion from Mitochondria to the Nucleus of Hydroxystyrylpyridinium by Introducing Only an Additional <i>o</i>-Hydroxyl Group

Aromatic cationic groups serve as crucial building blocks for the design of fluorescent probes targeting both the nucleus and mitochondria. Therefore, it is a significant challenge to develop aromatic cation-based probes that accurately image the nucleus without interference from mitochondria. However, this also presents an opportunity for rational design by modifying probes originally targeting mitochondria to redirect their targeting toward the nucleus. This study showcases the rapid development of a novel nucleus-targeting probe (DHSP) through a targeted conversion strategy based on structure modification of hydroxystyrylpyridinium (HSP), a well-established two-photon fluorescent probe that targets mitochondria. Importantly, DHSP, which is derived exclusively from introducing only an additional <i>o</i>-hydroxyl group into HSP, exhibits robust DNA-binding capability comparable to a commercially available nuclear dye 4',6-diamidino-2-phenylindole (DAPI). As a result, it rapidly enters the nucleus within 5 min and finds successful application in two-photon cellular and intravital imaging of the nucleus.