Detail of Fig2_onephoton_mito_fixed_SPIM
Project
SSBD:Repository
Title
Fluorescence images of mitochondria in fixed HeLa cells in the imaging mode of SPIM for single-photon activation
Description
The fluorescence images of mitochondria labeled with Skylan-NS in fixed HeLa cells were taken and reconstructed in the imaging mode of selective plane illumination microscopy (SPIM). The SPIM mode reduced the out-of-focus emission, whereas the lateral spatial resolution did not improve. Skylan-NS is a photoswitchable green fluorescent protein. CW laser is used for single-photon activation, which is more efficient and allows measurement with higher temporal resolution.
Release, Updated
2026-07-07
License
CC BY 4.0
Kind
Image data
File Formats
tif
Data size
9.0 MB
Datatype
-
Molecular Function (MF)
Biological Process (BP)
Cellular Component (CC)
mitochondrion
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
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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