扫描电镜
荧光团
活体细胞成像
显微镜
荧光
线粒体
细胞器
荧光显微镜
生物物理学
受激发射
超微结构
电子显微镜
荧光寿命成像显微镜
化学
细胞生物学
生物
光学
细胞
激光器
物理
解剖
生物化学
作者
Chenguang Wang,Masayasu Taki,Yoshikatsu Sato,Yasushi Tamura,Hideyuki Yaginuma,Yasushi Okada,Shigehiro Yamaguchi
标识
DOI:10.1073/pnas.1905924116
摘要
Stimulation emission depletion (STED) microscopy enables ultrastructural imaging of organelle dynamics with a high spatiotemporal resolution in living cells. For the visualization of the mitochondrial 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 mitochondrial cristae with a resolution of ∼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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