Detail of Fig5B_Mitochondrial_MitoPB



Project
Title
Time-lapse STED imaging of mitochondrial dynamics in MitoPB Yellow-labeled cells
Description
Time-lapse STED imaging of mitochondrial dynamics in MitoPB Yellow-labeled cells
Release, Updated
2022-11-23
License
CC-BY
Kind
Image data
File Formats
uncompressed TIFF
Data size
200.1 MB

Organism
Homo sapiens ( NCBITaxon:9606 )
Strain(s)
-
Cell Line
HeLa cell ( CLO_0003684 )
Reporter
MitoPB Yellow

Datatype
-
Molecular Function (MF)
Biological Process (BP)
mitochondrial fusion ( GO:0008053 )
Cellular Component (CC)
mitochondrion ( GO:0005739 ) mitochondrial crista ( GO:0030061 ) mitochondrial inner membrane ( GO:0005743 ) mitochondrial outer membrane ( GO:0005741 ) mitochondrial membrane ( GO:0031966 )
Biological Imaging Method
time lapse microscopy ( Fbbi:00000249 )
Stimulation emission depletion ( Fbbi:00000334 )
X scale
0.0227272 micrometer/pixel
Y scale
0.0227272 micrometer/pixel
Z scale
1 micrometer/slice
T scale
1.31 sec per time interval

Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-

Summary of Methods
See details in Wang C, et. al. (2019) Proc Natl Acad Sci U S A., 116(32):15817-15822.
Related paper(s)

Chenguang Wang, Masayasu Taki, Yoshikatsu Sato, Yasushi Tamura, Hideyuki Yaginuma, Yasushi Okada, Shigehiro Yamaguchi (2019) A photostable fluorescent marker for the superresolution live imaging of the dynamic structure of the mitochondrial cristae., Proceedings of the National Academy of Sciences of the United States of America, Volume 116, Number 32, pp. 15817-15822

Published in 2019 Aug 6 (Electronic publication in July 23, 2019, midnight )

(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 approximately 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.
(MeSH Terms)

Contact
Taki Masayasu, Yasushi Okada, Shigehiro Yamaguchi , Nagoya University, RIKEN, Nagoya University , Institute of Transformative Bio-Molecules, Center for Biosystems Dynamics Research , Institute of Transformative Bio-Molecules , Functional Organic Materials Laboratory, Laboratory for Cell Polarity Regulation , Functional Organic Materials Laboratory
Contributors

OMERO Dataset
OMERO Project
Source