Positive-Type Reversibly Photoswitching Red Fluorescent Protein for Dual-Color Superresolution Imaging with Single Light Exposure for Off-Switching

Ozaki-Noma, Ryohei; Wazawa, Tetsuichi; Kakizuka, Taishi et al.

ACS Nano, 2025, 19(7), 7188-7201

Number of Access:651（2025-11-02 16:29 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/101031

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Title
Positive-Type Reversibly Photoswitching Red Fluorescent Protein for Dual-Color Superresolution Imaging with Single Light Exposure for Off-Switching
Creator
CreatorName
Ozaki-Noma, Ryohei
NRID
1000080359851 1000080359851
researchmap permalink
wazawa wazawa
CreatorName
Wazawa, Tetsuichi
ORCID
0000-0003-4269-5923 0000-0003-4269-5923
researchmap permalink
t-kakizuka t-kakizuka
CreatorName
Kakizuka, Taishi
NRID
1000000812736 1000000812736
CreatorName
Shidara, Hisashi
NRID
1000080466432 1000080466432
CreatorName
Takemoto, Kiwamu
NRID
1000020311350 1000020311350
ORCID
0000-0003-2650-9895 0000-0003-2650-9895
researchmap permalink
ng1 ng1
Scopus Author ID
7403433377 7403433377
CreatorName
Nagai, Takeharu
Subject
chromophore phenolate
fluorescence
multiple equilibria
nanoscopy
positive-type reversibly photoswitching fluorescent protein
red fluorescent protein
superresolution imaging
Description
Ozaki-Noma R., Wazawa T., Kakizuka T., et al. Positive-Type Reversibly Photoswitching Red Fluorescent Protein for Dual-Color Superresolution Imaging with Single Light Exposure for Off-Switching. ACS Nano 19, 7188 (2025); https://doi.org/10.1021/acsnano.4c16847.
Abstract
Positive-type reversibly photoswitching fluorescent proteins (p-rsFPs) transition to a bright on-state upon light exposure for fluorescence excitation and to a dark off-state under a different wavelength. p-rsFPs are widely used in superresolution (SR) imaging techniques, offering simplified observation procedure and enhanced biocompatibility. Although some green p-rsFPs possess adequate photoproperties for SR imaging, all red p-rsFPs (p-rsRFPs) to date exhibit suboptimal properties, limiting the color palette for multiplexed SR imaging. Here, we present a p-rsRFP, rsZACRO, with 3.0-fold brighter fluorescence, 5.3-fold faster off-switching, and 1.5-fold higher on/off contrast than rsCherry, a conventional representative p-rsRFP. Using rsZACRO with superresolution polarization demodulation/on-state polarization angle narrowing (SPoD-OnSPAN), we successfully demonstrated SR imaging in the red spectrum and dual-color SR imaging with a single light for off-switching, visualizing vimentin intermediate filaments and actin filaments at higher spatial resolution than the diffraction limit of light in a living mammalian cell.
Publisher
American Chemical Society
Source Title
ACS Nano
Volume (Issue)
19(7)
Page
7188-7201
Date of Issued
2025-02-25
Language
English
Handle URL
https://hdl.handle.net/11094/101031
PISSN
19360851
eISSN
1936086X
Relation.isIdenticalTo
DOI (Publisher Version)
https://doi.org/10.1021/acsnano.4c16847
Access Rights
open access
CopyRight
This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
oaire:version
Version of Record
Category
学術雑誌論文 Journal Article