Simultaneous Zn2+ tracking in multiple organelles using super-resolution morphology-correlated organelle identification in living cells

Hongbao Fang, Shanshan Geng, Mingang Hao, Qixin Chen, Minglun Liu, Chunyan Liu, Zhiqi Tian, Chengjun Wang, Takanori Takebe, Jun-Lin Guan, Yuncong Chen (), Zijian Guo, Weijiang He () and Jiajie Diao ()
Additional contact information
Hongbao Fang: Nanjing University
Shanshan Geng: Nanjing University
Mingang Hao: University of Cincinnati College of Medicine
Qixin Chen: University of Cincinnati College of Medicine
Minglun Liu: Nanjing University
Chunyan Liu: Cincinnati Children’s Hospital Medical Center
Zhiqi Tian: University of Cincinnati College of Medicine
Chengjun Wang: Sinopec Shengli Petroleum Engineering Limited Company
Takanori Takebe: Cincinnati Children’s Hospital Medical Center
Jun-Lin Guan: University of Cincinnati College of Medicine
Yuncong Chen: Nanjing University
Zijian Guo: Nanjing University
Weijiang He: Nanjing University
Jiajie Diao: University of Cincinnati College of Medicine

Nature Communications, 2021, vol. 12, issue 1, 1-14

Abstract: Abstract Zn2+ plays important roles in metabolism and signaling regulation. Subcellular Zn2+ compartmentalization is essential for organelle functions and cell biology, but there is currently no method to determine Zn2+ signaling relationships among more than two different organelles with one probe. Here, we report simultaneous Zn2+ tracking in multiple organelles (Zn-STIMO), a method that uses structured illumination microscopy (SIM) and a single Zn2+ fluorescent probe, allowing super-resolution morphology-correlated organelle identification in living cells. To guarantee SIM imaging quality for organelle identification, we develop a new turn-on Zn2+ fluorescent probe, NapBu-BPEA, by regulating the lipophilicity of naphthalimide-derived Zn2+ probes to make it accumulate in multiple organelles except the nucleus. Zn-STIMO with this probe shows that CCCP-induced mitophagy in HeLa cells is associated with labile Zn2+ enhancement. Therefore, direct organelle identification supported by SIM imaging makes Zn-STIMO a reliable method to determine labile Zn2+ dynamics in various organelles with one probe. Finally, SIM imaging of pluripotent stem cell-derived organoids with NapBu-BPEA demonstrates the potential of super-resolution morphology-correlated organelle identification to track biospecies and events in specific organelles within organoids.

Date: 2021
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DOI: 10.1038/s41467-020-20309-7

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