High-throughput fluorescence lifetime imaging flow cytometry
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 unaff...
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| Format: | Article |
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Nature Portfolio
2024-09-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-51125-y |
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| author | 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 Keisuke Goda |
| author_facet | 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 Keisuke Goda |
| author_sort | Hiroshi Kanno |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-9e141a3a3e5140768e9b58bd2c11814e |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-9e141a3a3e5140768e9b58bd2c11814e2025-01-12T12:29:36ZengNature PortfolioNature Communications2041-17232024-09-0115111510.1038/s41467-024-51125-yHigh-throughput fluorescence lifetime imaging flow cytometryHiroshi Kanno0Kotaro Hiramatsu1Hideharu Mikami2Atsushi Nakayashiki3Shota Yamashita4Arata Nagai5Kohki Okabe6Fan Li7Fei Yin8Keita Tominaga9Omer Faruk Bicer10Ryohei Noma11Bahareh Kiani12Olga Efa13Martin Büscher14Tetsuichi Wazawa15Masahiro Sonoshita16Hirofumi Shintaku17Takeharu Nagai18Sigurd Braun19Jessica P. Houston20Sherif Rashad21Kuniyasu Niizuma22Keisuke Goda23Department of Chemistry, The University of TokyoDepartment of Chemistry, The University of TokyoDepartment of Chemistry, The University of TokyoDepartment of Neurosurgery, Tohoku University Graduate School of MedicineDepartment of Neurosurgery, Tohoku University Graduate School of MedicineDepartment of Neurosurgery, Tohoku University Graduate School of MedicineGraduate School of Pharmaceutical Sciences, The University of TokyoDepartment of Chemistry, The University of TokyoDepartment of Neurosurgical Engineering and Translational Neuroscience, Tohoku University Graduate School of MedicineDepartment of Neurosurgery, Tohoku University Graduate School of MedicineDepartment of Chemistry, The University of TokyoSANKEN (The Institute of Scientific and Industrial Research), Osaka UniversityMiltenyi Biotec B.V. & Co. KGMiltenyi Biotec B.V. & Co. KGMiltenyi Biotec B.V. & Co. KGSANKEN (The Institute of Scientific and Industrial Research), Osaka UniversityInstitute for Genetic Medicine, Hokkaido UniversityInstitute for Life and Medical Sciences, Kyoto UniversitySANKEN (The Institute of Scientific and Industrial Research), Osaka UniversityInstitute for Genetics, Justus-Liebig-University GiessenDepartment of Chemical and Materials Engineering, New Mexico State UniversityDepartment of Neurosurgical Engineering and Translational Neuroscience, Tohoku University Graduate School of MedicineDepartment of Neurosurgical Engineering and Translational Neuroscience, Tohoku University Graduate School of MedicineDepartment of Chemistry, The University of TokyoAbstract 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.https://doi.org/10.1038/s41467-024-51125-y |
| spellingShingle | 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 Keisuke Goda High-throughput fluorescence lifetime imaging flow cytometry Nature Communications |
| title | High-throughput fluorescence lifetime imaging flow cytometry |
| title_full | High-throughput fluorescence lifetime imaging flow cytometry |
| title_fullStr | High-throughput fluorescence lifetime imaging flow cytometry |
| title_full_unstemmed | High-throughput fluorescence lifetime imaging flow cytometry |
| title_short | High-throughput fluorescence lifetime imaging flow cytometry |
| title_sort | high throughput fluorescence lifetime imaging flow cytometry |
| url | https://doi.org/10.1038/s41467-024-51125-y |
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