Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 116, Issue 32, Pages 15817-15822Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1905924116
Keywords
superresolution; STED microscopy; fluorescence probe; mitochondrial cristae; live-cell imaging
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Funding
- JST PRESTO [JPMJPR16F5]
- JST CREST [JPMJCR15G2]
- JSPS KAKENHI [16H05119, 17H19511, 18K14721, JP16H06280]
- Grants-in-Aid for Scientific Research [16H05119, 18K14721] Funding Source: KAKEN
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Stimulation emission depletion (STED) microscopy enables ultrastructural imaging of organelle dynamics with a high spatiotemporal resolution in living cells. For the visualization of the mitochondria! membrane dynamics in STED microscopy, rationally designed mitochondrial fluorescent markers with enhanced photostability are required. Herein, we report the development of a superphotostable fluorescent labeling reagent with long fluorescence lifetime, whose design is based on a structurally reinforced naphthophosphole fluorophore that is conjugated with an electron-donating diphenylamino group. The combination of long-lived fluorescence and superphotostable features of the fluorophore allowed us to selectively capture the ultrastructures of the mitochondria! cristae with a resolution of similar to 60 nm when depleted at 660 nm. This chemical tool provides morphological information of the cristae, which has so far only been observed in fixed cells using electron microscopy. Moreover, this method gives information about the dynamic ultrastructures such as the intermembrane fusion in different mitochondria as well as the intercristae mergence in a single mitochondrion during the apoptosis-like mitochondrial swelling process.
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