Detail of Fig3C_3color_BY2_unmixed_images

(Too many images for preview; see images in SSBD:OMERO Dataset)

Time-lapse images of 3-color labeled tobacco BY-2 cell divisions during mitosis.
Time-lapse images of 3-color labeled tobacco BY-2 cell divisions during mitosis.
Release, Updated
Image data
File Formats
Data size
905.3 MB

Nicotiana tabacum ( NCBITaxon:4097 )
Cell Line

Molecular Function (MF)
Biological Process (BP)
cell division ( GO:0051301 ) mitotic cell cycle ( GO:0000278 )
Cellular Component (CC)
nucleus ( GO:0005634 )
Biological Imaging Method
time lapse microscopy ( Fbbi:00000249 )
X scale
0.23 micrometer/pixel
Y scale
0.23 micrometer/pixel
Z scale
0.5 micrometer/slice
T scale
60 seconds 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 Kamada, T, et. al. (2022) Sci Rep 12, 809
Related paper(s)

Takafumi Kamada, Kohei Otomo, Takashi Murata, Kaito Nakata, Shota Hiruma, Ryota Uehara, Mitsuyasu Hasebe, Tomomi Nemoto (2022) Low-invasive 5D visualization of mitotic progression by two-photon excitation spinning-disk confocal microscopy., Scientific reports, Volume 12, Number 1, pp. 809

Published in 2022 Jan 17 (Electronic publication in Jan. 17, 2022, midnight )

(Abstract) Non-linear microscopy, such as multi-photon excitation microscopy, offers spatial localities of excitations, thereby achieving 3D cross-sectional imaging with low phototoxicity even in thick biological specimens. We had developed a multi-point scanning two-photon excitation microscopy system using a spinning-disk confocal scanning unit. However, its severe color cross-talk has precluded multi-color simultaneous imaging. Therefore, in this study, we introduced a mechanical switching system to select either of two NIR laser light pulses and an image-splitting detection system for 3- or 4-color imaging. As a proof of concept, we performed multi-color fluorescent imaging of actively dividing human HeLa cells and tobacco BY-2 cells. We found that the proposed microscopy system enabled time-lapse multi-color 3D imaging of cell divisions while avoiding photodamage. Moreover, the application of a linear unmixing method to the 5D dataset enabled the precise separation of individual intracellular components in multi-color images. We thus demonstrated the versatility of our new microscopy system in capturing the dynamic processes of cellular components that could have multitudes of application.
(MeSH Terms)

Kohei Otomo, Tomomi Nemoto , Juntendo University, National Institutes of Natural Sciences , Graduate School of Medicine, Exploratory Research Center on Life and Living Systems

OMERO Dataset
OMERO Project