Raman microscopy of cryofixed biological specimens for high-resolution and high-sensitivity chemical imaging

Mizushima, Kenta; Kumamoto, Yasuaki; Tamura, Shoko et al.

Science Advances, 2024, 10(50)

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Identifier to cite or link to this item: https://hdl.handle.net/11094/100600

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Title
Raman microscopy of cryofixed biological specimens for high-resolution and high-sensitivity chemical imaging
Creator
CreatorName
Mizushima, Kenta
NRID
1000030611727 1000030611727
ORCID
0000-0002-4398-288X 0000-0002-4398-288X
researchmap permalink
jemappellekuma jemappellekuma
Scopus Author ID
24376935000 24376935000
CreatorName
Kumamoto, Yasuaki
CreatorName
Tamura, Shoko
NRID
1000090648221 1000090648221
researchmap permalink
masahito_yamanaka masahito_yamanaka
Scopus Author ID
26642849100 26642849100
CreatorName
Yamanaka, Masahito
NRID
1000050868768 1000050868768
CreatorName
Mochizuki, Kentaro
CreatorName
Li, Menglu
NRID
1000060755932 1000060755932
ORCID
0000-0001-7389-9342 0000-0001-7389-9342
researchmap permalink
s.egoshi s.egoshi
CreatorName
Egoshi, Syusuke
NRID
1000020415243 1000020415243
CreatorName
Dodo, Kosuke
NRID
1000010381956 1000010381956
CreatorName
Harada, Yoshinori
CreatorName
Smith, I. Nicholas
NRID
1000060192142 1000060192142
CreatorName
Sodeoka, Mikiko
NRID
1000060236619 1000060236619
CreatorName
Tanaka, Hideo
NRID
1000080362664 1000080362664
ORCID
0000-0002-2284-375X 0000-0002-2284-375X
researchmap permalink
read0089911 read0089911
Scopus Author ID
7404058829 7404058829
CreatorName
Fujita, Katsumasa
Description
Mizushima K., Kumamoto Y., Tamura S., et al. Raman microscopy of cryofixed biological specimens for high-resolution and high-sensitivity chemical imaging. Science Advances 10, (2024); https://doi.org/10.1126/sciadv.adn0110.
Abstract
Raman microscopy is an emerging molecular imaging technology, yet its signal-to- noise ratio (SNR) in measurements of biological specimens is severely limited because of the small cross section of Raman scattering. Here, we present Raman imaging techniques of cryofixed specimens to overcome SNR limitations by enabling long exposure of specimens under highly stabilized low-temperature conditions. The observation of frozen specimens in a cryostat at a constant low temperature immediately after rapid freezing enabled the improvement of SNR and enhanced the spatial and spectral resolution. We also confirmed that the cryofixation can preserve physicochemical states of specimens by observing alkyne-labeled coenzyme Q in cytosol and hemeproteins in acute ischemic myocardium, which cannot be done by fixation using chemical reagents. Last, we applied the technique for multiplex Raman imaging of label-free endogenous molecules and alkyne-tagged molecules in cryofixed HeLa cells, demonstrating its capability of high-content imaging of complex biological phenomena while maintaining physiological conditions.
Publisher
American Association for the Advancement of Science
Source Title
Science Advances
Volume (Issue)
10(50)
Date of Issued
2024-12-13
Language
English
Handle URL
https://hdl.handle.net/11094/100600
eISSN
23752548
Relation.isIdenticalTo
DOI (Publisher Version)
https://doi.org/10.1126/sciadv.adn0110
Access Rights
open access
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This article is licensed under a Creative Commons Attribution 4.0 International License.
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Category
学術雑誌論文 Journal Article