Time-deterministic cryo-optical microscopy

Tsuji, Kosuke; Yamanaka, Masahito; Kumamoto, Yasuaki; Tamura, Shoko; Miyamura, Wakana; Kubo, Toshiki; Mizushima, Kenta; Kono, Kakeru; Hirano, Hanae; Shiozaki, Momoko; Zhao, Xiaowei; Xi, Heqi; Sugiura, Kazunori; Fukushima, Shun-ichi; Kunimoto, Takumi; Tanabe, Yoshino; Nishida, Kentaro; Mochizuki, Kentaro; Harada, Yoshinori; Smith, Nicholas I.; Heintzmann, Rainer; Yu, Zhiheng; Wang, Meng C.; Nagai, Takeharu; Tanaka, Hideo; Fujita, Katsumasa

doi:https://doi.org/10.1038/s41377-025-01941-8

Time-deterministic cryo-optical microscopy

Tsuji, Kosuke; Yamanaka, Masahito; Kumamoto, Yasuaki et al.

Light: Science and Applications, 2025, 14, 275

アクセス数:276件（2026-02-15 20:10 集計）

固定URL: https://hdl.handle.net/11094/103490

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タイトル	Time-deterministic cryo-optical microscopy
著者	ORCID 0009-0006-4437-7485 0009-0006-4437-7485 Scopus Author ID 58632598000 58632598000 著者名 Tsuji, Kosuke
	NRID 1000090648221 1000090648221 ORCID 0000-0002-5862-177X 0000-0002-5862-177X researchmap permalink masahito_yamanaka masahito_yamanaka Scopus Author ID 26642849100 26642849100 著者名 Yamanaka, Masahito
	NRID 1000030611727 1000030611727 ORCID 0000-0002-4398-288X 0000-0002-4398-288X researchmap permalink jemappellekuma jemappellekuma Scopus Author ID 24376935000 24376935000 著者名 Kumamoto, Yasuaki
	NRID 1000030849261 1000030849261 著者名 Tamura, Shoko
	著者名 Miyamura, Wakana
	NRID 1000050975159 1000050975159 ORCID 0000-0002-5696-4111 0000-0002-5696-4111 researchmap permalink kubo_toshiki_8 kubo_toshiki_8 著者名 Kubo, Toshiki
	著者名 Mizushima, Kenta
	著者名 Kono, Kakeru
	著者名 Hirano, Hanae
	著者名 Shiozaki, Momoko
	著者名 Zhao, Xiaowei
	著者名 Xi, Heqi
	著者名 Sugiura, Kazunori
	NRID 1000000793720 1000000793720 著者名 Fukushima, Shun-ichi
	著者名 Kunimoto, Takumi
	著者名 Tanabe, Yoshino
	著者名 Nishida, Kentaro
	NRID 1000050868768 1000050868768 著者名 Mochizuki, Kentaro
	NRID 1000010381956 1000010381956 著者名 Harada, Yoshinori
	NRID 1000010362592 1000010362592 ORCID 0000-0002-7835-199X 0000-0002-7835-199X researchmap permalink read0089935 read0089935 Scopus Author ID 55262815400 55262815400 著者名 Smith, Nicholas I.
	著者名 Heintzmann, Rainer
	著者名 Yu, Zhiheng
	著者名 Wang, Meng C.
	NRID 1000020311350 1000020311350 ORCID 0000-0003-2650-9895 0000-0003-2650-9895 researchmap permalink ng1 ng1 Scopus Author ID 7403433377 7403433377 著者名 Nagai, Takeharu
	NRID 1000060236619 1000060236619 著者名 Tanaka, Hideo
	NRID 1000080362664 1000080362664 ORCID 0000-0002-2284-375X 0000-0002-2284-375X researchmap permalink read0089911 read0089911 Scopus Author ID 7404058829 7404058829 著者名 Fujita, Katsumasa
内容	Tsuji K., Yamanaka M., Kumamoto Y., et al. Time-deterministic cryo-optical microscopy. Light: Science and Applications 14, 275 (2025); https://doi.org/10.1038/s41377-025-01941-8.
抄録	Fluorescence microscopy enables the visualization of cellular morphology, molecular distribution, ion distribution, and their dynamic behaviors during biological processes. Enhancing the signal-to-noise ratio (SNR) in fluorescence imaging improves the quantification accuracy and spatial resolution; however, achieving high SNR at fast image acquisition rates, which is often required to observe cellular dynamics, still remains a challenge. In this study, we developed a technique to rapidly freeze biological cells in milliseconds during optical microscopy observation. Compared to chemical fixation, rapid freezing provides rapid immobilization of samples while more effectively preserving the morphology and conditions of cells. This technique combines the advantages of both live-cell and cryofixation microscopy, i.e., temporal dynamics and high SNR snapshots of selected moments, and is demonstrated by fluorescence and Raman microscopy with high spatial resolution and quantification under low temperature conditions. Furthermore, we also demonstrated that intracellular calcium dynamics can be frozen rapidly and visualized using fluorescent ion indicators, suggesting that ion distribution and conformation of the probe molecules can be fixed both spatially and temporally. These results confirmed that our technique can time-deterministically suspend and visualize cellular dynamics while preserving molecular and ionic states, indicating the potential to provide detailed insights into sample dynamics with improved spatial resolution and temporal accuracy in observations.
出版者	Springer Nature
収録物名	Light: Science and Applications
巻(号)	14
ページ	275
刊行年月	2025-08-23
言語	英語
ハンドルURL	https://hdl.handle.net/11094/103490
PISSN	20955545
eISSN	20477538
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1038/s41377-025-01941-8
アクセス権	open access
権利情報	This article is licensed under a Creative Commons Attribution 4.0 International License.
出版タイプ	Version of Record
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCMI資源タイプ	text
DCTERMS.bibliographicCitation	Light: Science and Applications. 2025, 14, p. 275
DC.title	Time-deterministic cryo-optical microscopy
DC.creator	Tsuji, Kosuke
	Yamanaka, Masahito
	Kumamoto, Yasuaki
	Tamura, Shoko
	Miyamura, Wakana
	Kubo, Toshiki
	Mizushima, Kenta
	Kono, Kakeru
	Hirano, Hanae
	Shiozaki, Momoko
	Zhao, Xiaowei
	Xi, Heqi
	Sugiura, Kazunori
	Fukushima, Shun-ichi
	Kunimoto, Takumi
	Tanabe, Yoshino
	Nishida, Kentaro
	Mochizuki, Kentaro
	Harada, Yoshinori
	Smith, Nicholas I.
	Heintzmann, Rainer
	Yu, Zhiheng
	Wang, Meng C.
	Nagai, Takeharu
	Tanaka, Hideo
	Fujita, Katsumasa
DC.publisher	Springer Nature
DC.language' scheme='DCTERMS.RFC1766	英語
DC.type' scheme='DCTERMS.DCMIType	text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2025-08-23
DC.identifier	https://doi.org/10.1038/s41377-025-01941-8
DC.description	Tsuji K., Yamanaka M., Kumamoto Y., et al. Time-deterministic cryo-optical microscopy. Light: Science and Applications 14, 275 (2025); https://doi.org/10.1038/s41377-025-01941-8.
DCTERMS.abstract	Fluorescence microscopy enables the visualization of cellular morphology, molecular distribution, ion distribution, and their dynamic behaviors during biological processes. Enhancing the signal-to-noise ratio (SNR) in fluorescence imaging improves the quantification accuracy and spatial resolution; however, achieving high SNR at fast image acquisition rates, which is often required to observe cellular dynamics, still remains a challenge. In this study, we developed a technique to rapidly freeze biological cells in milliseconds during optical microscopy observation. Compared to chemical fixation, rapid freezing provides rapid immobilization of samples while more effectively preserving the morphology and conditions of cells. This technique combines the advantages of both live-cell and cryofixation microscopy, i.e., temporal dynamics and high SNR snapshots of selected moments, and is demonstrated by fluorescence and Raman microscopy with high spatial resolution and quantification under low temperature conditions. Furthermore, we also demonstrated that intracellular calcium dynamics can be frozen rapidly and visualized using fluorescent ion indicators, suggesting that ion distribution and conformation of the probe molecules can be fixed both spatially and temporally. These results confirmed that our technique can time-deterministically suspend and visualize cellular dynamics while preserving molecular and ionic states, indicating the potential to provide detailed insights into sample dynamics with improved spatial resolution and temporal accuracy in observations.
DC.rights	This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	103490
citation_title	Time-deterministic cryo-optical microscopy
citation_author	Tsuji, Kosuke
	Yamanaka, Masahito
	Kumamoto, Yasuaki
	Tamura, Shoko
	Miyamura, Wakana
	Kubo, Toshiki
	Mizushima, Kenta
	Kono, Kakeru
	Hirano, Hanae
	Shiozaki, Momoko
	Zhao, Xiaowei
	Xi, Heqi
	Sugiura, Kazunori
	Fukushima, Shun-ichi
	Kunimoto, Takumi
	Tanabe, Yoshino
	Nishida, Kentaro
	Mochizuki, Kentaro
	Harada, Yoshinori
	Smith, Nicholas I.
	Heintzmann, Rainer
	Yu, Zhiheng
	Wang, Meng C.
	Nagai, Takeharu
	Tanaka, Hideo
	Fujita, Katsumasa
citation_publisher	Springer Nature
citation_language	英語
citation_date	2025-08-23
citation_journal_title	Light: Science and Applications
citation_volume	14
citation_firstpage	275
citation_doi	https://doi.org/10.1038/s41377-025-01941-8
citation_public_url	https://hdl.handle.net/11094/103490

Time-deterministic cryo-optical microscopy

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