Intracellular oxygen imaging using small ratiometric oxygen probes

• Project/Area Number: 26702011
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Biomedical engineering/Biomaterial science and engineering
• Research Institution: Gunma University
• Principal Investigator: YOSHIHARA Toshitada 群馬大学, 理工学研究科, 准教授 (10375561)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥23,920,000 (Direct Cost: ¥18,400,000、Indirect Cost: ¥5,520,000); Fiscal Year 2016: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2015: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000); Fiscal Year 2014: ¥11,440,000 (Direct Cost: ¥8,800,000、Indirect Cost: ¥2,640,000)
• Keywords: りん光 / レシオ / イリジウム錯体 / 酸素 / 蛍光 / 顕微鏡 / 細胞 / 呼吸 / 細胞・組織 / 分析化学 / イメージング

Outline of Final Research Achievements

We designed and synthesized ratiometric molecular probes consisting of a blue fluorescent coumarin and a red phosphorescent cationic iridium complex connected by oligo proline of oligo arginine linkers, respectively, for real time sensing of oxygen levels in living cells. The introduction of oligo arginine linkers improved dramatically the cellular uptake efficiency compared to that of oligo proline linkers. We succeed the detection of oxygen levels in living cells using our small ratiometric oxygen probes.

Research Products

• [Journal Article] Oxygen Imaging of Living Cells and Tissues Using Luminescent Molecular Probes (2017)
  • Author(s): T. Yoshihara, Y. Hirakawa, M. Hosaka, M. Nangaku and S. Tobita
  • Journal Title: J. Photochem. Photobiol. C: Photochem. Rev. Volume: 30 Pages: 71-95
  • DOI: 10.1016/j.jphotochemrev.2017.01.001
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Interrogation of Metabolic and Oxygen States of Tumors with Fiber-Based Luminescence Lifetime Spectroscopy (2017)
  • Author(s): M. Lukina, A. Orlova, M. Shirmanova, D. Shirokov, A. Pavlikov, A. Neubauer, H. Studier, W. Becker, E. Zagaynova, T. Yoshihara, S. Tobita and V. Shcheslavskiy
  • Journal Title: Opt. Lett. Volume: 42 Issue: 4 Pages: 731-734
  • DOI: 10.1364/ol.42.000731
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Intracellular and in vivo oxygen sensing using phosphorescent iridium(III) complexes (2016)
  • Author(s): Tobita S, Yoshihara T
  • Journal Title: Current Opinion in Chemical Biology Volume: 33 Pages: 39-45
  • DOI: 10.1016/j.cbpa.2016.05.017
  • NAID: 130005163791
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Biological oxygen sensing via two-photon absorption by an Ir(III) complex using a femtosecond fiber laser (2016)
  • Author(s): Morimoto H, Fujii A, Suzuki Y, Yoshihara T, Tobita S, Kawamata J
  • Journal Title: Japanese Journal of Applied Physics Volume: 55 Issue: 9 Pages: 092401-092403
  • DOI: 10.7567/jjap.55.092401
  • NAID: 210000147062
  • Peer Reviewed
• [Journal Article] Phosphorescence quenching of neutral and cationic iridium(III) complexes by molecular oxygen and aromatic electron acceptors (2016)
  • Author(s): N. Hasebe, Y. Deguchi, S. Murayama, T. Yoshihara, H. Horiuchi, T. Okutsu, S. Tobita
  • Journal Title: J. Photochem. Photobiol. A Volume: 324 Pages: 134-144
  • DOI: 10.1016/j.jphotochem.2016.03.008
  • Peer Reviewed
• [Journal Article] Ratiometric molecular probes based on dual emission of a blue fluorescent coumarin and a red phosphorescent cationic iridium(III) complex for intracellular oxygen sensing (2015)
  • Author(s): Yoshihara T, Murayama S, Tobita S
  • Journal Title: Sensors Volume: 15 Issue: 6 Pages: 13503-13521
  • DOI: 10.3390/s150613503
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Quantitating intracellular oxygen tension in vivo by phosphorescence lifetime measurement (2015)
  • Author(s): Hirakawa Y, Yoshihara T, Kamiya M, Mimura I, Fujikura D, Masuda T, Kikuchi R, Takahashi I, Urano Y, Tobita S, Nangaku M.
  • Journal Title: Sci Rep Volume: 5 Issue: 1 Pages: 17838-17838
  • DOI: 10.1038/srep17838
  • Peer Reviewed / Open Access
• [Journal Article] Intracellular and in vivo oxygen sensing using phosphorescent Ir(III) complexes with a modified acetylacetonato ligand (2015)
  • Author(s): Yoshihara T, Hosaka M, Terata M, Ichikawa K, Murayama S, Tanaka A, Mori M, Itabashi H, Takeuchi T, Tobita S.
  • Journal Title: Anal. Chem. Volume: 87 Issue: 5 Pages: 2710-2717
  • DOI: 10.1021/ac5040067
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Mitochondria-targeted oxygen probes based on cationic iridium complexes with a 5-amino-1,10-phenanthroline ligand (2015)
  • Author(s): Yoshihara T, Murayama S, Masuda T, Kikuchi T, Yoshida K, Hosaka M, Tobita S.
  • Journal Title: J. Photochem. Photobiol. A: Chem Volume: 299 Pages: 172-182
  • DOI: 10.1016/j.jphotochem.2014.11.004
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Intracellular and in vivo Oxygen Sensing by Using Phosphorescence Lifetime Methods (2016)
  • Author(s): T. Yoshihara and S. Tobita
  • Organizer: Frontiers 2016 Symposium
  • Place of Presentation: Lausanne (Switzerland)
  • Int'l Joint Research / Invited
• [Presentation] 水溶性イリジウム錯体を用いた生体内酸素濃度計測用発光プローブの開発 (2016)
  • Author(s): 綿貫千優，吉原利忠，飛田成史
  • Organizer: 日本化学会関東支部群馬地区研究交流発表会
  • Place of Presentation: 高崎量子応用研究所（群馬県・高崎市）
• [Presentation] 共焦点PLIM法による細胞および細胞スフェロイドの酸素濃度計測 (2016)
  • Author(s): 水上輝市，吉原利忠，飛田成史
  • Organizer: 日本化学会関東支部群馬地区研究交流発表会
  • Place of Presentation: 高崎量子応用研究所（群馬県・高崎市）
• [Presentation] 細胞内移行性を高めたレシオ型生体酸素プローブ (2016)
  • Author(s): 山崎莉穂，安ヵ川真美，吉原利忠，飛田成史
  • Organizer: 第6回CSJ化学フェスタ
  • Place of Presentation: タワーホテル船堀（東京都・江戸川区）
• [Presentation] cRGDペプチドを結合させた腫瘍集積性イリジウム錯体の合成と生体酸素プローブへの応用 (2016)
  • Author(s): 中村俊介，吉原利忠，穂坂正博，飛田成史
  • Organizer: 第14回がんとハイポキシア研究会
  • Place of Presentation: 岐阜グランドホテル（岐阜県・岐阜市）
• [Presentation] 細胞膜透過性を高めたレシオ酸素プローブの開発および細胞内酸素濃度計測 (2016)
  • Author(s): 安ヵ川真美，吉原利忠，飛田成史
  • Organizer: 第14回がんとハイポキシア研究会
  • Place of Presentation: 岐阜グランドホテル（岐阜県・岐阜市）
• [Presentation] オルガネラ特異的イリジウム錯体を用いた細胞内酸素濃度測定 (2016)
  • Author(s): 平野翔太，高橋一平，吉原利忠，飛田成史
  • Organizer: 第10回分子科学討論会
  • Place of Presentation: 神戸ファッションマート（兵庫県・神戸市）
• [Presentation] 顕微りん光寿命計測に基づく細胞・組織内酸素イメージング (2016)
  • Author(s): 高橋一平，水上輝市，平野翔太，穂坂正博，吉原利忠，飛田成史
  • Organizer: 2016年光化学討論会
  • Place of Presentation: 東京大学駒場第一キャンパス（東京都・目黒区）
• [Presentation] Oxygen Concentration Measurements of Cell Spheroids by Means of Confocal Phosphorescence Lifetime Imaging Microscopy (2016)
  • Author(s): Kiichi Muzukami，Toshitada Yoshihara，Seiji Tobita
  • Organizer: 第11回日本分子イメージング学会学術大会
  • Place of Presentation: 神戸国際会議場（兵庫県・神戸市）
• [Presentation] Developments of Ratiometric Oxygen Probes for Intracellular Oxygen Sensing (2016)
  • Author(s): Toshitada Yoshihara，Saori Murayama，Seiji Tobita
  • Organizer: 第11回日本分子イメージング学会学術大会
  • Place of Presentation: 神戸国際会議場（兵庫県・神戸市）
• [Presentation] Ratiometric Optical Sensors Based on Dual Emission of a Blue Fluorescent Coumarin and a Red Phosphorescent Cationic Iridium(III) Complex for Oxygen Sensing in Living Cells (2015)
  • Author(s): T. Yoshihara, S. Murayama, and S. Tobita
  • Organizer: The International Chemical Congress of Pacific Basin Societies 2015
  • Place of Presentation: Honolulu (U. S. A.)
  • Year and Date: 2015-12-17
  • Int'l Joint Research
• [Presentation] Ratiometric Optical Sensors for Intracellular Oxygen Sensing (2015)
  • Author(s): T. Yoshihara, S. Murayama, Y. Yamaguchi, and S. Tobita
  • Organizer: Third International Symposium on the Photofunctional Chemistry of Complex Systems
  • Place of Presentation: Maui (U. S. A.)
  • Year and Date: 2015-12-13
  • Int'l Joint Research / Invited
• [Presentation] Two Dimensional Oxygen Imaging in Living Cells by Using a Gated ICCD Camera (2015)
  • Author(s): T. Yoshihara, I. Takahashi, and S. Tobita
  • Organizer: The Fifth International Symposium on Element Innovation
  • Place of Presentation: 桐生市市民文化会館(群馬県・桐生市）
  • Year and Date: 2015-11-30
  • Int'l Joint Research
• [Presentation] Tuning Emission Wavelengths of Metal Complexes Operative in Chemical Biology (2015)
  • Author(s): T. Yoshihara, M. Hosaka, T. Takeuchi, and S. Tobita
  • Organizer: 錯体化学会第65回討論会
  • Place of Presentation: 奈良女子大学（奈良県・奈良市）
  • Year and Date: 2015-09-21
  • Invited
• [Presentation] レシオ型分子酸素計の光物理特性および細胞内酸素センシング (2015)
  • Author(s): 吉原利忠，村山沙織，飛田成史
  • Organizer: 2015年光化学討論会
  • Place of Presentation: 大阪市立大学（大阪府・大阪市）
  • Year and Date: 2015-09-11
• [Presentation] In Vivo Oxygen Sensing Using Phosphorescence Lifetime Measurements of Cationic Iridium(III) Complexes (2015)
  • Author(s): T. Yoshihara, M. Hosaka, M. Terata, K. Ichikawa, M. Mori, H. Itabashi, T. Takeuchi, and S. Tobita
  • Organizer: The 3rd Conference of the Japanese Association for Hypoxia Biology
  • Place of Presentation: 早稲田大学先端生命医科学(東京都・新宿区)
  • Year and Date: 2015-07-25
  • Int'l Joint Research
• [Presentation] イリジウム錯体のりん光寿命計測による細胞・組織内の酸素濃度定量 (2015)
  • Author(s): 吉原利忠，穂坂正博，寺田幹，市川和貴，森勝伸，板橋英之，竹内利行，飛田成史
  • Organizer: 第10回日本分子イメージング学会学術大会
  • Place of Presentation: タワーホール船堀(東京都・江戸川区)
  • Year and Date: 2015-05-20
• [Presentation] りん光性イリジウム錯体を用いた細胞・組織内酸素レベル計測 (2015)
  • Author(s): 吉原利忠
  • Organizer: 第54回日本生体医工学会
  • Place of Presentation: 名古屋国際会議場（愛知県・名古屋市）
  • Year and Date: 2015-05-08
  • Invited
• [Presentation] カチオン性イリジウム錯体のりん光寿命測定による細胞内酸素レベル計測 (2015)
  • Author(s): 吉原利忠，増田剛，森勝伸，板橋英之，飛田成史
  • Organizer: 日本化学会第95春季年会(2015)
  • Place of Presentation: 日本大学理工学部（千葉県船橋市）
  • Year and Date: 2015-03-27
• [Presentation] Quantitative Analysis of Oxygen in Living Cells and Tissues Using Iridium(III) Complexes with Cationic Substituent (2014)
  • Author(s): T. Yoshihara, M. Hosaka, K. Kishimoto, H. Akiyama, and S. Tobita
  • Organizer: 1st International Symposium of Gunma University Medical Innovation and 6th International Conference on Advanced Micro-Device Engineering
  • Place of Presentation: Kiryu, Japan
  • Year and Date: 2014-12-05
• [Presentation] Developments of Mitochondria-Specific Oxygen Probes Based on Cationic Iridium(III) Complexes (2014)
  • Author(s): T. Yoshihara, S. Murayama, T. Kikuchi, and S. Tobita
  • Organizer: The Fourth International Symposium on Element Innovation
  • Place of Presentation: Kiryu, Japan
  • Year and Date: 2014-11-24
• [Presentation] イリジウム錯体のりん光を利用した細胞・組織内酸素濃度定量法の開発 (2014)
  • Author(s): 吉原利忠，飛田成史
  • Organizer: 生物発光化学発光研究会第31回学術講演会
  • Place of Presentation: 信州大学繊維学部（長野県上田市）
  • Year and Date: 2014-11-01
  • Invited
• [Presentation] ミトコンドリア局在性を示すカチオン性イリジウム錯体の光物理特性および細胞内挙動 (2014)
  • Author(s): 吉原利忠，村山沙織，菊池俊毅，飛田成史
  • Organizer: 2014年光化学討論会
  • Place of Presentation: 北海道大学（北海道札幌市）
  • Year and Date: 2014-10-11
• [Presentation] 細胞内移行性を向上させたレシオ型酸素プローブ分子の開発 (2014)
  • Author(s): 吉原利忠，村山沙織，飛田成史
  • Organizer: 第8回分子科学総合討論会2014東広島
  • Place of Presentation: 広島大学（広島県東広島市）
  • Year and Date: 2014-09-22
• [Presentation] Phosphorescent Molecular Probes for Imaging of Oxygen Levels in Living Cells and Hypoxic Tissues (2014)
  • Author(s): T. Yoshihara
  • Organizer: China/Japan Young Chemists Forum: Molecular Imaging for Chemical Biology
  • Place of Presentation: Beijing, China
  • Year and Date: 2014-08-05
  • Invited
• [Presentation] ミトコンドリア局在性を示すカチオン性イリジウム錯体を用いた生細胞イメージング (2014)
  • Author(s): 吉原利忠，村山沙織，菊池俊毅，竹内利行，飛田成史
  • Organizer: 第2回低酸素研究会
  • Place of Presentation: 早稲田大学先端生命医科学センター（東京都新宿区）
  • Year and Date: 2014-07-26
• [Presentation] Photophysical Properties of Cationic Iridium Complexes with Mitochondria Specifity and Their Intracellular Behavior (2014)
  • Author(s): T. Yoshihara, S. Murayama, T. Kikuchi, T. Takeuchi, and S. Tobita
  • Organizer: The XXVth IUPAC Symposium on Photochemistry
  • Place of Presentation: Bordeaux, France
  • Year and Date: 2014-07-14
• [Presentation] ミトコンドリア局在性を示すカチオン性イリジウム錯体の開発および生細胞イメージング (2014)
  • Author(s): 吉原利忠，村山沙織，菊池俊毅，飛田成史
  • Organizer: 第9回日本分子イメージング学会学術大会
  • Place of Presentation: 千里ライフサイエンスセンター(大阪府豊中市）
  • Year and Date: 2014-05-23
• [Book] DOJIN BIOSCIENCE SERIES 22 がんの分子イメージング (2015)
  • Author(s): 吉原利忠，飛田成史
  • Total Pages: 269
  • Publisher: 化学同人
• [Remarks] 群馬大学理工学府分子科学部門飛田研究室
  • URL: http://tobita-lab.chem-bio.st.gunma-u.ac.jp/
• [Remarks] 群馬大学理工学府分子科学部門飛田研究室ホームページ
  • URL: http://tobita-lab.chem-bio.st.gunma-u.ac.jp/
• [Remarks]
  • URL: http://tobita-lab.chem-bio.st.gunma-u.ac.jp/
• [Patent(Industrial Property Rights)] レシオ法を用いた酸素濃度測定試薬 (2016)
  • Inventor(s): 吉原利忠，安ヵ川真美，飛田成史
  • Industrial Property Rights Holder: 国立大学法人群馬大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2016-205799
  • Filing Date: 2016-10-20