Mitochondrial dynamics quantitatively revealed by STED nanoscopy with an enhanced squaraine variant probe

Xusan Yang (), Zhigang Yang (), Zhaoyang Wu, Ying He, Chunyan Shan, Peiyuan Chai, Chenshuo Ma, Mi Tian, Junlin Teng, Dayong Jin, Wei Yan, Pintu Das, Junle Qu () and Peng Xi ()
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Xusan Yang: Peking University
Zhigang Yang: Shenzhen University
Zhaoyang Wu: Peking University
Ying He: Shenzhen University
Chunyan Shan: Peking University
Peiyuan Chai: Peking University
Chenshuo Ma: Rutgers University
Mi Tian: Shenzhen University
Junlin Teng: Peking University
Dayong Jin: Southern University of Science and Technology
Wei Yan: Shenzhen University
Pintu Das: Shenzhen University
Junle Qu: Shenzhen University
Peng Xi: Peking University

Nature Communications, 2020, vol. 11, issue 1, 1-9

Abstract: Abstract Mitochondria play a critical role in generating energy to support the entire lifecycle of biological cells, yet it is still unclear how their morphological structures evolve to regulate their functionality. Conventional fluorescence microscopy can only provide ~300 nm resolution, which is insufficient to visualize mitochondrial cristae. Here, we developed an enhanced squaraine variant dye (MitoESq-635) to study the dynamic structures of mitochondrial cristae in live cells with a superresolution technique. The low saturation intensity and high photostability of MitoESq-635 make it ideal for long-term, high-resolution (stimulated emission depletion) STED nanoscopy. We performed time-lapse imaging of the mitochondrial inner membrane over 50 min (3.9 s per frame, with 71.5 s dark recovery) in living HeLa cells with a resolution of 35.2 nm. The forms of the cristae during mitochondrial fusion and fission can be clearly observed. Our study demonstrates the emerging capability of optical STED nanoscopy to investigate intracellular physiological processes with nanoscale resolution for an extended period of time.

Date: 2020
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DOI: 10.1038/s41467-020-17546-1

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