Detail of ExtFig6b_mClover3
Project
SSBD:Repository
Title
Time-lapse images of CAP-H-mClover3 in HCT116 cells during prometaphase
Description
Time-lapse images of CAP-H-mClover3 in HCT116 cells during prometaphase. The cells were volume imaged (z-step, 0.25 micrometer; z-range, 2.5 micrometer) every 6.9 seconds. With CRISPR-Cas9 genome-editing technique, a subunit of condensin I (CAP-H) was endogenously tagged at the C terminus with mClover3 (CAP-H-mClover3) in the HCT116 cells.
Release, Updated
2025-11-26
License
CC BY 4.0
Kind
Image data
File Formats
.tif
Data size
12.5 MB
Datatype
-
Molecular Function (MF)
regulate the chromosome assembly and segregation
Biological Process (BP)
chromosome organization
Cellular Component (CC)
nucleus
chromosome
Biological Imaging Method
spinning disk confocal microscopy
(
Fbbi:00000253
)
single-beam laser-scanning confocal microscopy (LSCM)
single-beam laser-scanning confocal microscopy (LSCM)
X scale
-
Y scale
-
Z scale
-
T scale
6.9 seconds
Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
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Objective model
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Filter set
-
Summary of Methods
Ando R, Shimozono S, Ago H, Takagi M, Sugiyama M, Kurokawa H, Hirano M, Niino Y, Ueno G, Ishidate F, Fujiwara T, Okada Y, Yamamoto M, Miyawaki A. StayGold variants for molecular fusion and membrane-targeting applications. Nat Methods. 2024 Apr;21(4):648-656.
Related paper(s)
Ryoko Ando, Satoshi Shimozono, Hideo Ago, Masatoshi Takagi, Mayu Sugiyama, Hiroshi Kurokawa, Masahiko Hirano, Yusuke Niino, Go Ueno, Fumiyoshi Ishidate, Takahiro Fujiwara, Yasushi Okada, Masaki Yamamoto, Atsushi Miyawaki (2023) StayGold variants for molecular fusion and membrane-targeting applications., Nature methods
Published in 2023 Nov 30 (Electronic publication in Nov. 30, 2023, midnight )
- doi: 10.1038/s41592-023-02085-6
-
pmid: 38036853
(Abstract) Although StayGold is a bright and highly photostable fluorescent protein, its propensity for obligate dimer formation may hinder applications in molecular fusion and membrane targeting. To attain monovalent as well as bright and photostable labeling, we engineered tandem dimers of StayGold to promote dispersibility. On the basis of the crystal structure of this fluorescent protein, we disrupted the dimerization to generate a monomeric variant that offers improved photostability and brightness compared to StayGold. We applied the new monovalent StayGold tools to live-cell imaging experiments using spinning-disk laser-scanning confocal microscopy or structured illumination microscopy. We achieved cell-wide, high-spatiotemporal resolution and sustained imaging of dynamic subcellular events, including the targeting of endogenous condensin I to mitotic chromosomes, the movement of the Golgi apparatus and its membranous derivatives along microtubule networks, the distribution of cortical filamentous actin and the remolding of cristae membranes within mobile mitochondria.
Contact
Atsushi Miyawaki
, RIKEN
, Laboratory for Cell Function Dynamics, RIKEN Center for Brain Science
, Laboratory for Cell Function Dynamics, RIKEN Center for Brain Science
Contributors
OMERO Project