Detail of Fig3_twophoton_Lifeact_SPASIM_2D
Project
SSBD:Repository
Title
The x-y image of actin filaments of living HeLa cells in 3DSIM mode for two-photon activation
Description
An x-y image of actin filaments in living HeLa cells visualized with rsGamillus-S was obtained using selective-plane-activation structured illumination microscopy (SPA-SIM) with two-photon activation and a scanned Bessel beam. 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
18.0 MB
Datatype
-
Molecular Function (MF)
Biological Process (BP)
Cellular Component (CC)
actin filament
Biological Imaging Method
structured illumination microscopy (FBbi_00000332)
X scale
70.9 micrometer
Y scale
70.9 micrometer
Z scale
-
T scale
-
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
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
-
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