1. An assembly-regulated SNAP-tag fluorogenic probe for long-term super-resolution imaging of mitochondrial dynamics.
- Author
-
Liu, Wenjuan, Qiao, Qinglong, Zheng, Jiazhu, Chen, Jie, Zhou, Wei, Xu, Ning, Li, Jin, Miao, Lu, and Xu, Zhaochao
- Subjects
- *
HIGH resolution imaging , *MITOCHONDRIAL membranes , *FLUORESCENCE microscopy , *INTRAMOLECULAR charge transfer , *MITOCHONDRIA , *MEMBRANE potential , *MEMBRANE proteins - Abstract
Super-resolution fluorescence microscopy has emerged as a powerful tool for studying mitochondrial dynamics in living cells. However, the lack of photostable and chemstable probe makes long-term super-resolution imaging of mitochondria still a challenging work. Herein, we reported a 4-azetidinyl-naphthliamide derived SNAP-tag probe AN-BG exhibiting excellent fluorogenicity and photostability for long-term super-resolution imaging of mitochondrial dynamics. The azetidinyl group and naphthalimide fluorophore are in a flat conformation which can effectively suppress twisted intramolecular charge transfer and then effectively improve the brightness and photostability. This planarized molecular structure is conducive to the formation of fluorescence-quenched J -aggregates, and the protein labeling process will depolymerize the probes and restore fluorescence. Fluorescent labeling mitochondrial inner membrane proteins via SNAP tags overcomes the shortcomings that variations in mitochondrial inner membrane potential will release probes attached to mitochondria by electrostatic interactions. Therefore, AN-BG realized the stable labeling of mitochondria and the long-term imaging of mitochondrial dynamics under super-resolution microscopy. Image 1 • A naphthliamide-derived fluorogenic probe for SNAP-tag was developed. • Fluorescent labeling of mitochondria was achieved via SNAP-tag fused proteins. • Long-term super-resolution imaging of mitochondrial dynamics was achieved. • The dynamic mitochondrial fusion and fission were imaged with fluorescent nanoscopy. • Two-color SIM imaging revealed multiple modes of action between mitochondria and lysosomes. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF