Detail of FigS12_twophoton_nuclei_rsG_SPASIM
Project
SSBD:Repository
Title
Fluorescence images of nuclei labeled by rsGamillus-S in HeLa cell spheroids in SPA-SIM mode for two-photon activation
Description
Fluorescence images of nuclei in HeLa cell spheroids expressing rsGamillus-S-labeled histone H2B were obtained using selective-plane-activation structured illumination microscopy (SPA-SIM) with two-photon activation by a Bessel sheet. rsGamillus-S is an acid-resistance photoswitchable fluorescent protein, which owns exceptionally high on-switching rate to show the nuclei more clearly. Two-photon activation using a scanned Bessel beam produces a wider FOV and a higher axial resolution.
Release, Updated
2026-07-07
License
CC BY 4.0
Kind
Image data
File Formats
tif
Data size
36.0 MB
Datatype
-
Molecular Function (MF)
Biological Process (BP)
Cellular Component (CC)
nucleus
Biological Imaging Method
structured illumination microscopy (FBbi_00000332)
X scale
141.8 micrometer
Y scale
141.8 micrometer
Z scale
-
T scale
-
Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
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Microscope model
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Detector model
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Objective model
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Filter set
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Summary of Methods
Temma K, Oketani R, Kubo T, Bando K, Maeda S, Sugiura K, Matsuda T, Heintzmann R, Kaminishi T, Fukuda K, Hamasaki M, Nagai T, Fujita K. Selective-plane-activation structured illumination microscopy. Nat Methods. 2024 May;21(5):889-896.
Related paper(s)
Kenta Temma, Ryosuke Oketani, Toshiki Kubo, Kazuki Bando, Shunsuke Maeda, Kazunori Sugiura, Tomoki Matsuda, Rainer Heintzmann, Tatsuya Kaminishi, Koki Fukuda, Maho Hamasaki, Takeharu Nagai, Katsumasa Fujita (2024) Selective-plane-activation structured illumination microscopy., Nature methods
Published in 2024 Apr 5 (Electronic publication in April 5, 2024, midnight )
- doi: 10.1038/s41592-024-02236-3
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pmid: 38580844
(Abstract) The background light from out-of-focus planes hinders resolution enhancement in structured illumination microscopy when observing volumetric samples. Here we used selective plane illumination and reversibly photoswitchable fluorescent proteins to realize structured illumination within the focal plane and eliminate the out-of-focus background. Theoretical investigation of the imaging properties and experimental demonstrations show that selective plane activation is beneficial for imaging dense microstructures in cells and cell spheroids.
Contact
Katsumasa Fujita
, Osaka University
, Department of Applied Physics
, Department of Applied Physics
Contributors
OMERO Project
Source