Summary of 242-Wazawa-CellDyn

SSBD:database
SSBD:database URL
Title
Time-lapse super resolution imaging of Kohinoor2.0 or Kohinoor in live cells.
Description
-
Relase date
2022-11-23
Updated date
-
License
CC BY
Kind
Image data based on Experiment
Number of Datasets
16 ( Image datasets: 16, Quantitative data datasets: 0 )
Size of Datasets
2.8 GB ( Image datasets: 2.8 GB, Quantitative data datasets: 0 bytes )

Organism(s)
Homo sapiens, Cercopithecidae
Cell lines(s)
HeLa cell, COS7 cell
Gene symbol(s)
NA
Protein name(s)
NA

Datatype
-
Molecular Function (MF)
Biological Process (BP)
Actin filament dynamics, mitochondrial fusion, mitochondrial fission, Photoswitching of Kohinoor 2.0, Photobleaching of Kohinoor 2.0
Cellular Component (CC)
actin filament, Vimentin intermediate filament, mitochondrial matrix
Biological Imaging Method
time lapse microscopy, time lapse super-resolution microscopy
X scale
26 nanometer per pixel, 156 nanometer per pixel, 20 nanometer/pixel
Y scale
26 nanometer per pixel, 156 nanometer per pixel, 20 nanometer/pixel
Z scale
-
T scale
3 minutes per frame, 16.7 milisecond per frame, 1 hour per frame, 167 millisecond per frame, 0.5 second per frame

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

Related paper(s)

Tetsuichi Wazawa, Ryohei Noma, Shusaku Uto, Kazunori Sugiura, Takashi Washio, Takeharu Nagai (2021) A photoswitchable fluorescent protein for hours-time-lapse and sub-second-resolved super-resolution imaging., Microscopy (Oxford, England), Volume 70, Number 4, pp. 340-352

Published in 2021 Aug 9

(Abstract) Reversibly photoswitchable fluorescent proteins (RSFPs) are a class of fluorescent proteins whose fluorescence can be turned on and off by light irradiation. RSFPs have become essential tools for super-resolution (SR) imaging. Because most SR imaging techniques require high-power-density illumination, mitigating phototoxicity in cells due to intense light irradiation has been a challenge. Although we previously developed an RSFP named Kohinoor to achieve SR imaging with low phototoxicity, the photoproperties were insufficient to move a step further to explore the cellular dynamics by SR imaging. Here, we show an improved version of RSFP, Kohinoor2.0, which is suitable for SR imaging of cellular processes. Kohinoor2.0 shows a 2.6-fold higher fluorescence intensity, 2.5-fold faster chromophore maturation and 1.5-fold faster off-switching than Kohinoor. The analysis of the pH dependence of the visible absorption band revealed that Kohinoor2.0 and Kohinoor were in equilibria among multiple fluorescently bright and dark states, with the mutations introduced into Kohinoor2.0 bringing about a higher stabilization of the fluorescently bright states compared to Kohinoor. Using Kohinoor2.0 with our SR imaging technique, super-resolution polarization demodulation/on-state polarization angle narrowing, we conducted 4-h time-lapse SR imaging of an actin filament network in mammalian cells with a total acquisition time of 480 s without a noticeable indication of phototoxicity. Furthermore, we demonstrated the SR imaging of mitochondria dynamics at a time resolution of 0.5 s, in which the fusion and fission processes were clearly visualized. Thus, Kohinoor2.0 is shown to be an invaluable RSFP for the SR imaging of cellular dynamics.
(MeSH Terms)

Contact
Takeharu Nagai , Osaka University , SANKEN , Nagai Laboratory
Contributors


Dataset List of 242-Wazawa-CellDyn

#
Dataset ID
Kind
Size
4D View
SSBD:OMERO
Download BDML
Download Images
# 8790
Dataset Kind Image data
Dataset Size 160.0 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8791
Dataset Kind Image data
Dataset Size 320.0 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8792
Dataset Kind Image data
Dataset Size 326.7 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8793
Dataset Kind Image data
Dataset Size 72.6 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8794
Dataset Kind Image data
Dataset Size 448.4 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8795
Dataset Kind Image data
Dataset Size 99.6 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8796
Dataset Kind Image data
Dataset Size 7.0 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8797
Dataset Kind Image data
Dataset Size 8.0 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8798
Dataset Kind Image data
Dataset Size 5.0 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8799
Dataset Kind Image data
Dataset Size 7.0 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8800
Dataset Kind Image data
Dataset Size 7.0 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8801
Dataset Kind Image data
Dataset Size 7.0 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8802
Dataset Kind Image data
Dataset Size 7.0 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8803
Dataset Kind Image data
Dataset Size 5.0 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8804
Dataset Kind Image data
Dataset Size 1000.4 MB
4D view
SSBD:OMERO
Download BDML
Download Image data

# 8805
Dataset Kind Image data
Dataset Size 372.1 MB
4D view
SSBD:OMERO
Download BDML
Download Image data