High-throughput fluorescence lifetime imaging flow cytometry

Hiroshi Kanno (), Kotaro Hiramatsu, Hideharu Mikami, Atsushi Nakayashiki, Shota Yamashita, Arata Nagai, Kohki Okabe, Fan Li, Fei Yin, Keita Tominaga, Omer Faruk Bicer, Ryohei Noma, Bahareh Kiani, Olga Efa, Martin Büscher, Tetsuichi Wazawa, Masahiro Sonoshita, Hirofumi Shintaku, Takeharu Nagai, Sigurd Braun, Jessica P. Houston, Sherif Rashad, Kuniyasu Niizuma and Keisuke Goda ()
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Hiroshi Kanno: The University of Tokyo
Kotaro Hiramatsu: The University of Tokyo
Hideharu Mikami: The University of Tokyo
Atsushi Nakayashiki: Tohoku University Graduate School of Medicine
Shota Yamashita: Tohoku University Graduate School of Medicine
Arata Nagai: Tohoku University Graduate School of Medicine
Kohki Okabe: The University of Tokyo
Fan Li: The University of Tokyo
Fei Yin: Tohoku University Graduate School of Medicine
Keita Tominaga: Tohoku University Graduate School of Medicine
Omer Faruk Bicer: The University of Tokyo
Ryohei Noma: Osaka University
Bahareh Kiani: Miltenyi Biotec B.V. & Co. KG
Olga Efa: Miltenyi Biotec B.V. & Co. KG
Martin Büscher: Miltenyi Biotec B.V. & Co. KG
Tetsuichi Wazawa: Osaka University
Masahiro Sonoshita: Hokkaido University
Hirofumi Shintaku: Kyoto University
Takeharu Nagai: Osaka University
Sigurd Braun: Justus-Liebig-University Giessen
Jessica P. Houston: New Mexico State University
Sherif Rashad: Tohoku University Graduate School of Medicine
Kuniyasu Niizuma: Tohoku University Graduate School of Medicine
Keisuke Goda: The University of Tokyo

Nature Communications, 2024, vol. 15, issue 1, 1-15

Abstract: Abstract Flow cytometry is a vital tool in biomedical research and laboratory medicine. However, its accuracy is often compromised by undesired fluctuations in fluorescence intensity. While fluorescence lifetime imaging microscopy (FLIM) bypasses this challenge as fluorescence lifetime remains unaffected by such fluctuations, the full integration of FLIM into flow cytometry has yet to be demonstrated due to speed limitations. Here we overcome the speed limitations in FLIM, thereby enabling high-throughput FLIM flow cytometry at a high rate of over 10,000 cells per second. This is made possible by using dual intensity-modulated continuous-wave beam arrays with complementary modulation frequency pairs for fluorophore excitation and acquiring fluorescence lifetime images of rapidly flowing cells. Moreover, our FLIM system distinguishes subpopulations in male rat glioma and captures dynamic changes in the cell nucleus induced by an anti-cancer drug. FLIM flow cytometry significantly enhances cellular analysis capabilities, providing detailed insights into cellular functions, interactions, and environments.

Date: 2024
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DOI: 10.1038/s41467-024-51125-y

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