Detail of Fig2_onephoton_act_TL_SPIM


Project
SSBD:Repository
Title
Time-lapse images of actin filaments of living HeLa cells in SPIM mode for single-photon activation
Description
The actin filaments visualized by Skylan-NS in living HeLa cells were observed in selective plane illumination microscopy (SPIM) using single-photon activation mode. SPIM visualizes actin movement at a lower spatial resolution than that of SPA-SIM. 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
14.0 MB

Organism
Homo sapiens ( NCBI:txid9606 )
Strain(s)
-
Cell Line
HeLa-S3 cell

Datatype
-
Molecular Function (MF)
Biological Process (BP)
Cellular Component (CC)
actin filament
Biological Imaging Method
structured illumination microscopy (FBbi_00000332) time lapse microscopy (FBbi_00000249)
X scale
29.6 micrometer
Y scale
29.6 micrometer
Z scale
-
T scale
7 seconds

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

(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 Dataset
OMERO Project
Source