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High-throughput fluorescence lifetime imaging flow cytometry

Author

Listed:
  • Hiroshi Kanno

    (The University of Tokyo
    Tohoku University Graduate School of Medicine)

  • Kotaro Hiramatsu

    (The University of Tokyo
    Kyushu University)

  • Hideharu Mikami

    (The University of Tokyo
    Hokkaido University)

  • 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
    Tohoku University)

  • Kuniyasu Niizuma

    (Tohoku University Graduate School of Medicine
    Tohoku University Graduate School of Medicine
    Tohoku University)

  • Keisuke Goda

    (The University of Tokyo
    Wuhan University
    University of California)

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.

Suggested Citation

  • 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 Ki, 2024. "High-throughput fluorescence lifetime imaging flow cytometry," Nature Communications, Nature, vol. 15(1), pages 1-15, December.
    • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-51125-y
    DOI: 10.1038/s41467-024-51125-y
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    References listed on IDEAS

    as
    1. Hugh Sparks & Hiroshi Kondo & Steven Hooper & Ian Munro & Gordon Kennedy & Christopher Dunsby & Paul French & Erik Sahai, 2018. "Heterogeneity in tumor chromatin-doxorubicin binding revealed by in vivo fluorescence lifetime imaging confocal endomicroscopy," Nature Communications, Nature, vol. 9(1), pages 1-14, December.
    2. Natalia Ochocka & Pawel Segit & Kacper Adam Walentynowicz & Kamil Wojnicki & Salwador Cyranowski & Julian Swatler & Jakub Mieczkowski & Bozena Kaminska, 2021. "Single-cell RNA sequencing reveals functional heterogeneity of glioma-associated brain macrophages," Nature Communications, Nature, vol. 12(1), pages 1-14, December.
    3. Masako Nishikawa & Hiroshi Kanno & Yuqi Zhou & Ting-Hui Xiao & Takuma Suzuki & Yuma Ibayashi & Jeffrey Harmon & Shigekazu Takizawa & Kotaro Hiramatsu & Nao Nitta & Risako Kameyama & Walker Peterson & , 2021. "Massive image-based single-cell profiling reveals high levels of circulating platelet aggregates in patients with COVID-19," Nature Communications, Nature, vol. 12(1), pages 1-12, December.
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