Investigation into cell heterogeneity using ultra-fast imaging technique

• Project/Area Number: 15J02613
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 国内
• Research Field: Biomedical engineering/Biomaterial science and engineering
• Research Institution: The University of Tokyo
• Principal Investigator: 小林 博文 東京大学, 理学系研究科, 特別研究員(DC1)
• Project Period (FY): 2015-04-24 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥2,800,000 (Direct Cost: ¥2,800,000); Fiscal Year 2017: ¥900,000 (Direct Cost: ¥900,000); Fiscal Year 2016: ¥900,000 (Direct Cost: ¥900,000); Fiscal Year 2015: ¥1,000,000 (Direct Cost: ¥1,000,000)
• Keywords: ラベルフリー / 薬剤応答性 / 薬剤耐性 / 機械学習 / 全血 / 異種移植 / ハイスループット / ハイコンテント / 画像処理 / 高速イメージング / 蛍光イメージング / 形態学的変化 / 細胞刺激 / 周波数多重 / 画像計測

Outline of Annual Research Achievements

細胞の薬剤応答性は細胞の根本的な生物学を探るのに重要なだけでなく、新規薬剤の創造にも重要な要素となります。本研究では、薬剤で処理された細胞の無標識画像を撮り、薬剤に誘導された形態学的変化を画像から評価することにより、細胞の薬剤応答性を定量評価するという手法を提案しました。受け入れ研究室で開発されたOptofluitic time-stretch (OTS)顕微鏡により、大量の細胞の無標識画像が高いスループットで撮影できることから、従来のハイスループットスクリーニングと同じ速さで画像情報が得られるため、スクリーニングを初め、血液検査などへの応用も期待されます。
前年度までに、ヒト乳がん細胞株であるMCF-7やヒト白血病細胞株であるK562を用いた原理実証を果たし、無標識画像を用いて、薬剤によって形態変化が誘導されること、そしてこれが無標識画像によって検出・評価可能であることが分かりました。今年度はこれまでの原理実証をさらに深く、多方面から調べ、本研究で提案する手法の限界がどこにあるのか、どこまで実用性があるのかを明らかにしました。また、本手法の汎用性を調べるために、様々な試料を試しました。例えば、白血病細胞の薬剤応答性の違いから細胞の薬剤耐性を調べるられることが分かっただけでなく、薬剤処理した白血病細胞の薬剤応答性が、全血においても調べられることを示すことができました。この結果は、本研究で提案する手法が血液検査にも応用できることを示唆しています。さらにマウスに移植したヒトがん細胞からも、同様の手法で薬剤応答性が調べられることを示しました。これにより、本研究で開発、提案する手法が非常に幅広い応用性、汎用性を持つことが分かり、蛍光標識を使わない新たなスクリーニング技術及び血液検査技術を確立しました。

Research Progress Status

29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

29年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] Carnegie Mellon University(米国)
• [Int'l Joint Research] National Chiao Tung University(台湾)
• [Journal Article] Ultrafast confocal fluorescence microscopy beyond the fluorescence lifetime limit (2018)
  • Author(s): Mikami H, Harmon J, Kobayashi H, Hamad S, Wang Y, Iwata O, Suzuki K, Ito T, Aisaka Y, Kutsuna N, Nagasawa K, Watarai H, Ozeki Y, Goda K.
  • Journal Title: Optica Volume: 5 Issue: 2 Pages: 117-126
  • DOI: 10.1364/optica.5.000117
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Optofluidic time-stretch quantitative phase microscopy (2018)
  • Author(s): Guo Baoshan、Lei Cheng、Wu Yi、Kobayashi Hirofumi、Ito Takuro、Yalikun Yaxiaer、Lee Sangwook、Isozaki Akihiro、Li Ming、Jiang Yiyue、Yasumoto Atsushi、Di Carlo Dino、Tanaka Yo、Yatomi Yutaka、Ozeki Yasuyuki、Goda Keisuke
  • Journal Title: Methods Volume: 136 Pages: 116-125
  • DOI: 10.1016/j.ymeth.2017.10.004
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Label-free detection of cellular drug responses by high-throughput bright-field imaging and machine learning (2017)
  • Author(s): Kobayashi Hirofumi、Lei Cheng、Wu Yi、Mao Ailin、Jiang Yiyue、Guo Baoshan、Ozeki Yasuyuki、Goda Keisuke
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 12454-12454
  • DOI: 10.1038/s41598-017-12378-4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Label-free detection of aggregated platelets in blood by machine-learning-aided optofluidic time-stretch microscopy. (2017)
  • Author(s): Jiang Y, Lei C, Yasumoto A, Kobayashi H, Aisaka Y, Ito T, Guo B, Nitta N, Kutsuna N, Ozeki Y, Nakagawa A, Yatomi Y, Goda K.
  • Journal Title: Lab on a Chip Volume: 17 Issue: 14 Pages: 2426-2434
  • DOI: 10.1039/c7lc00396j
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] High-throughput, label-free, single-cell, microalgal lipid screening by machine-learning-equipped optofluidic time-stretch quantitative phase microscopy (2017)
  • Author(s): Guo Baoshan、Lei Cheng、Kobayashi Hirofumi、Ito Takuro、Yalikun Yaxiaer、Jiang Yiyue、Tanaka Yo、Ozeki Yasuyuki、Goda Keisuke
  • Journal Title: Cytometry Part A Volume: 91 Issue: 5 Pages: 494-502
  • DOI: 10.1002/cyto.a.23084
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] GHz Optical Time-Stretch Microscopy by Compressive Sensing (2017)
  • Author(s): Lei Cheng、Wu Yi、Sankaranarayanan Aswin C.、Chang Shih-Min、Guo Baoshan、Sasaki Naoto、Kobayashi Hirofumi、Sun Chia-Wei、Ozeki Yasuyuki、Goda Keisuke
  • Journal Title: IEEE Photonics Journal Volume: 9 Issue: 2 Pages: 1-8
  • DOI: 10.1109/jphot.2017.2676349
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] High-throughput label-free screening of euglena gracilis with optofluidic time-stretch quantitative phase microscopy、High-Speed Biomedical Imaging and Spectroscopy (2017)
  • Author(s): aoshan Guo ; Cheng Lei ; Takuro Ito ; Yalikun Yaxiaer ; Hirofumi Kobayashi ; Yiyue Jiang ; Yo Tanaka ; Yasuyuki Ozeki ; Keisuke Goda
  • Journal Title: Proc. SPIE 10076 Volume: 10076 Pages: 1-6
  • DOI: 10.1117/12.2251157
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Enhanced speed in fluorescence imaging using beat frequency multiplexing (2016)
  • Author(s): Hideharu Mikami ; Hirofumi Kobayashi ; Yisen Wang ; Syed Hamad ; Yasuyuki Ozeki ; Keisuke Goda
  • Journal Title: Proc. SPIE 9720, High-Speed Biomedical Imaging and Spectroscopy: Toward Big Data Instrumentation and Management Volume: 9720 Pages: 1-6
  • DOI: 10.1117/12.2211836
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] Label-free image-based detection of drug resistance with optofluidic time-stretch microscopy (2017)
  • Author(s): Hirofumi Kobayashi, Cheng Lei, Ailin Mao, Yiyue Jiang, Baoshan Guo, Yasuyuki Ozeki, Keisuke Goda
  • Organizer: Photonics West 2017
  • Place of Presentation: San Francisco (USA)
  • Year and Date: 2017-01-28
  • Int'l Joint Research
• [Presentation] Label-free image-based detection of drug resistance with optofluidic time-stretch microscopy (2017)
  • Author(s): Kobayashi, Hirofumi Lei, Cheng Mao, Ailin Jiang, Yiyue Guo, Baoshan Ozeki, Yasuyuki Goda, Keisuke
  • Organizer: Photonics West 2017
  • Int'l Joint Research
• [Presentation] High-throughput label-free detection of aggregate platelets with optofluidic time-stretch microscopy (2017)
  • Author(s): Jiang, Yiyue Lei, Cheng Yasumoto, Atsushi Ito, Takuro Guo, Baoshan Kobayashi, Hirofumi Ozeki, Yasuyuki Yatomi, Yutaka Goda, Keisuke
  • Organizer: Photonics West 2017
  • Int'l Joint Research
• [Presentation] High-throughput label-free screening of euglena gracilis with optofluidic time-stretch quantitative phase microscopy (2017)
  • Author(s): Guo, Baoshan Lei, Cheng Ito, Takuro Yaxiaer, Yalikun Kobayashi, Hirofumi Jiang, Yiyue Tanaka, Yo Ozeki, Yasuyuki Goda, Keisuke
  • Organizer: Photonics West 2017
  • Int'l Joint Research
• [Presentation] High-throughput, label-free, multivariate cell analysis with optofluidic time-stretch microscopy (2017)
  • Author(s): C. Lei, B. Guo, Y. Jiang, Y. Wu, H. Kobayashi, T Ito, A. Yasumoto, Y. Yatomi, Y. Ozeki, and K. Goda,
  • Organizer: Conference on Lasers and Electro-Optics (CLEO) Pacific Rim 2017
  • Int'l Joint Research
• [Presentation] Morphology-based cancer detection with optofluidic time-stretch microscopy (2016)
  • Author(s): Hirofumi Kobayashi, Cheng Lei, Ailin Mao, Yiyue Jiang, Baoshan Guo, Yasuyuki Ozeki, Keisuke Goda
  • Organizer: Photonics Asia 2016
  • Place of Presentation: Beijing (China)
  • Year and Date: 2016-10-12
  • Int'l Joint Research
• [Presentation] Enhanced speed in fluorescence imaging using beat frequency multiplexing (2016)
  • Author(s): Hideharu Mikami ; Hirofumi Kobayashi ; Yisen Wang ; Syed Hamad ; Yasuyuki Ozeki ; Keisuke Goda
  • Organizer: Photonics West 2016
  • Place of Presentation: San Francisco, California, United States
  • Year and Date: 2016-02-13
  • Int'l Joint Research