RRC ID 58312
Author Wang C, Taki M, Sato Y, Tamura Y, Yaginuma H, Okada Y, Yamaguchi S.
Title A photostable fluorescent marker for the superresolution live imaging of the dynamic structure of the mitochondrial cristae.
Journal Proc Natl Acad Sci U S A
Abstract 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.
Volume 116(32)
Pages 15817-15822
Published 2019-8-6
DOI 10.1073/pnas.1905924116
PII 1905924116
PMID 31337683
PMC PMC6689947
MeSH Cell Line Fluorescent Dyes / chemistry* Humans Imaging, Three-Dimensional* Light* Mitochondria / chemistry* Mitochondrial Membranes / metabolism Time-Lapse Imaging
IF 9.58
Times Cited 15
Resource
DNA material pN1-OPA1(123Nt)-Halo (RDB17747) pN1-TOMM20-Halo (RDB17750)