Detail of Fig4G_Resort
Project
Title
RESORT imaging of live HeLa cells stained with DAE620-Mito
Description
RESORT (reversible saturable optical Raman transitions) imaging of live HeLa cells stained with DAE620-Mito (triphenylphosphonium (Mito-tag) into DAE620 via a butyl linker).
Release, Updated
2024-11-25
License
CC BY
Kind
Image data
File Formats
.tif
Data size
492.2 KB
Datatype
-
Molecular Function (MF)
Biological Process (BP)
Cellular Component (CC)
mitochondrion
(
GO:0005739
)
Biological Imaging Method
inelastic scattering of photons
(
Fbbi:00000589
)
reversible saturable optical Raman transitions (RESORT) microscopy
reversible saturable optical Raman transitions (RESORT) microscopy
X scale
0.08 micrometer/pixel
Y scale
0.08 micrometer/pixel
Z scale
-
T scale
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Image Acquisition
Experiment type
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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
See details in Shou J, et. al. Sci Adv. 2023 Jun 16;9(24):eade9118.
Related paper(s)
Jingwen Shou, Ayumi Komazawa, Yuusaku Wachi, Minoru Kawatani, Hiroyoshi Fujioka, Spencer John Spratt, Takaha Mizuguchi, Kenichi Oguchi, Hikaru Akaboshi, Fumiaki Obata, Ryo Tachibana, Shun Yasunaga, Yoshio Mita, Yoshihiro Misawa, Ryosuke Kojima, Yasuteru Urano, Mako Kamiya, Yasuyuki Ozeki (2023) Super-resolution vibrational imaging based on photoswitchable Raman probe., Science advances, Volume 9, Number 24, pp. eade9118
Published in 2023 Jun 16 (Electronic publication in June 16, 2023, midnight )
- doi: 10.1126/sciadv.ade9118
-
pmid: 37327330
(Abstract) Super-resolution vibrational microscopy is promising to increase the degree of multiplexing of nanometer-scale biological imaging because of the narrower spectral linewidth of molecular vibration compared to fluorescence. However, current techniques of super-resolution vibrational microscopy suffer from various limitations including the need for cell fixation, high power loading, or complicated detection schemes. Here, we present reversible saturable optical Raman transitions (RESORT) microscopy, which overcomes these limitations by using photoswitchable stimulated Raman scattering (SRS). We first describe a bright photoswitchable Raman probe (DAE620) and validate its signal activation and depletion characteristics when exposed to low-power (microwatt level) continuous-wave laser light. By harnessing the SRS signal depletion of DAE620 through a donut-shaped beam, we demonstrate super-resolution vibrational imaging of mammalian cells with excellent chemical specificity and spatial resolution beyond the optical diffraction limit. Our results indicate RESORT microscopy to be an effective tool with high potential for multiplexed super-resolution imaging of live cells.(MeSH Terms)
Contact
Yasuyuki Ozeki, Mako Kamiya
, The University of Tokyo, The University of Tokyo
, Department of Electrical Engineering and Information Systems, Graduate School of Medicine
Contributors
Jingwen Shou, Ayumi Komazawa, Yuusaku Wachi, Hiroyoshi Fujioka, Minoru Kawatani, Takaha Mizuguchi, Hikaru Akaboshi
OMERO Project
Source