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论文类型： 期刊论文

发表时间： 2018-10-01

发表刊物： NATURE PROTOCOLS

收录刊物： PubMed、SCIE

卷号： 13

期号： 10

页面范围： 2348-2361

ISSN号： 1754-2189

摘要： Hypochlorous acid (HOCI) is a critical member of the reactive oxygen species (ROS) produced by immune cells to fight infections. On the other hand, HOCI in homeostasis causes oxidative damage to biomolecules and is linked to many diseases, including inflammatory, neurodegenerative, and cardiovascular diseases. Herein, we detail a procedure for the preparation of a boron-dipyrromethene (BODIPY)-derived fluorescent probe for HOCI (BCIO) and its application as an imaging reagent in living cells. BCIO is synthesized in one pot through a four-step procedure that is nearly the same as that for conventional BODIPY dye preparation, except for the ratio of starting materials. BCIO has an extremely rapid response (saturated within seconds) and is ultrasensitive to HOCI. The detection limit of BCIO reaches the subnanomolar range, which is the highest HOCI sensitivity to date. Taking advantage of the ultrasensitive character of BCIO, we have previously demonstrated its ability to detect endogenous HOCI generated by macrophages and shown that it can also be used to discriminate cancer cell lines (which show high HOCI production) from non-cancer cell lines (which show low HOCI production). The protocol requires similar to 2 d for probe synthesis and up to similar to 18 h for fluorescence imaging and flow cytometry assays.