Development of microscopic single-molecule pathological examination device for DNA methylation

• Project/Area Number: 26750146
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biomedical engineering/Biomaterial science and engineering
• Research Institution: Nagoya University
• Principal Investigator: ONOSHIMA Daisuke 名古屋大学, 未来社会創造機構, 特任講師 (40510219)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2016: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: １分子計測(SMD) / 細胞・組織 / マイクロ・ナノデバイス

Outline of Final Research Achievements

Image analysis of methylated DNA has a great potential for exploring epigenomic change in cancer cells. In order to visualize signal variation caused by DNA methylation, it is crucial that the DNA molecules are arrayed in parallel direction inside the narrow microscopic field. Here we show that methylated DNA molecules can be trapped and stretched at the patterned grooves on the surface of microfluidic channel by flow. The demonstrated DNA aligning technique enables simultaneous molecular imaging and detection for cancer diagnosis at the single molecule level.

Research Products

• [Journal Article] Cell deposition microchip with micropipette control over liquid interface motion (2018)
  • Author(s): D. Onoshima, Y. Hattori, H. Yukawa, K. Ishikawa, M. Hori, and Y. Baba
  • Journal Title: Cell Med Volume: -
  • Peer Reviewed / Open Access
• [Journal Article] Facile fabrication of a poly(ethylene terephthalate) membrane filter with precise arrangement of through-holes (2018)
  • Author(s): N. Kihara, H. Odaka, D. Kuboyama, D. Onoshima, K. Ishikawa, Y. Baba, and M. Hori
  • Journal Title: Jpn. J. Appl. Phys Volume: 57 Issue: 3 Pages: 037001-037001
  • DOI: 10.7567/jjap.57.037001
  • NAID: 210000148722
  • Peer Reviewed
• [Journal Article] Measurement of DNA Length Changes upon CpG Hypermethylation by Microfluidic Molecular Stretching (2017)
  • Author(s): D. Onoshima, N. Kawakita, D. Takeshita, H. Niioka, H. Yukawa, J. Miyake, and Y. Baba
  • Journal Title: Cell Med. Volume: 9 Issue: 1-2 Pages: 61-66
  • DOI: 10.3727/215517916x693087
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Microfluidic DNA stretching device for single-molecule diagnostics (2016)
  • Author(s): D. Onoshima and Y. Baba
  • Journal Title: Methods Mol. Biol. Volume: 1547 Pages: 105-111
  • DOI: 10.1007/978-1-4939-6734-6_8
  • ISBN: 9781493967322, 9781493967346
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Multifunctional nanobiodevices in medical sciences (2015)
  • Author(s): D. Onoshima, H. Yukawa, and Y. Baba
  • Journal Title: Adv. Drug Deliv. Rev. Volume: 95 Pages: 1-1
  • DOI: 10.1016/j.addr.2015.11.006
  • Peer Reviewed
• [Journal Article] Multifunctional quantum dots-based cancer diagnostics and stem cell therapeutics for regenerative medicine (2015)
  • Author(s): D. Onoshima, H. Yukawa, and Y. Baba
  • Journal Title: Adv. Drug Deliv. Rev. Volume: 95 Pages: 2-14
  • DOI: 10.1016/j.addr.2015.08.004
  • Peer Reviewed
• [Journal Article] Superhydrophilic Glass Membrane Device with Open-Microhole Array for Filtering and Counting Rare Tumor Cells (2015)
  • Author(s): A. Yonese, D. Onoshima, H. Yukawa, K. Ishikawa, M. Hori, and Y. Baba
  • Journal Title: Micro Total Analysis Systems Volume: 1 Pages: 493-495
  • Peer Reviewed
• [Journal Article] Microfluidic transfer of liquid interface for parallel stretching and stamping of terminal-unmodified single DNA molecules in zigzag-shaped microgrooves (2014)
  • Author(s): H. Yasaki, D. Onoshima, T. Yasui, H. Yukawa, N. Kaji, and Y. Baba
  • Journal Title: Lab Chip Volume: 15 Issue: 1 Pages: 135-140
  • DOI: 10.1039/c4lc00990h
  • Peer Reviewed
• [Journal Article] Novel positively charged nanoparticle labeling for in vivo imaging of adipose tissue-derived stem cells. (2014)
  • Author(s): Yukawa H, Nakagawa S, Yoshizumi Y, Watanabe M, Saito H, Miyamoto Y, Noguchi H, Oishi K, Ono K, Sawada M, Kato I, Onoshima D, Obayashi M, Hayashi Y, Kaji N, Ishikawa T, Hayashi S, Baba Y.
  • Journal Title: PLoS One. Volume: 9 Issue: 11 Pages: e110142-e110142
  • DOI: 10.1371/journal.pone.0110142
  • Peer Reviewed / Open Access
• [Journal Article] On-chip analysis of intermittent molecular encounters in nuclease digestion of specific DNA sequence (2014)
  • Author(s): D. Onoshima, N. Kaji, M. Tokeshi, and Y. Baba
  • Journal Title: Biophys. J Volume: 103 Issue: 2 Pages: 699a-700a
  • DOI: 10.1016/j.bpj.2013.11.3869
  • Peer Reviewed
• [Journal Article] Microfluidic Stretching of DNA with fluorescent gold nanoparticle for optical/electron microscopic imaging of a single DNA methylation (2014)
  • Author(s): D. Takeshita, D. Onoshima, H. Yukawa, T. Yasui, N. Kaji, and Y. Baba
  • Journal Title: Micro Total Analysis Systems Volume: 1 Pages: 2348-2350
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Sizing and sorting of single DNA molecules by microfluidic molecular combing device (2014)
  • Author(s): D. Onoshima, N. Kawakita, D. Takeshita, H. Yukawa, and Y. Baba
  • Journal Title: Micro Total Analysis Systems Volume: 1 Pages: 1775-1777
  • Peer Reviewed
• [Journal Article] ここまで進んだがんの診断・血中マーカーによる診断と呼気診断の最前線 (2014)
  • Author(s): 小野島大介、馬場嘉信
  • Journal Title: ライフライン21 がんの先進医療 Volume: 15 Pages: 46-48
• [Presentation] Cell deposition and isolation with micropipette control over liquid interface motion in microfluidic channel (2017)
  • Author(s): D. Onoshima, H. Yukawa, Y. Hattori, K. Ishikawa, M. Hori, and Y. Baba
  • Organizer: The 21st International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2017)
  • Int'l Joint Research
• [Presentation] A novel membrane separation technology for cancer cells in blood (2017)
  • Author(s): D. Onoshima
  • Organizer: International Symposium on Pure & Applied Chemistry (ISPAC) 2017
  • Int'l Joint Research / Invited
• [Presentation] Applications of bioimaging devices for cell transplantation and tumorigenic analysis in regenerative medicine (2016)
  • Author(s): D. Onoshima
  • Organizer: International Symposium on Pure & Applied Chemistry (ISPAC) 2016
  • Place of Presentation: Borneo Convention Center Kuching, Kuching, Sarawak, Malaysia
  • Year and Date: 2016-08-15
  • Int'l Joint Research / Invited
• [Presentation] ポータブル・メディカル・デバイスが実現する未来社会 (2016)
  • Author(s): 小野島大介
  • Organizer: 第１２回プラズマ医療サイエンスの扉
  • Place of Presentation: 名古屋大学VBLベンチャーホール3F
  • Year and Date: 2016-06-17
  • Invited
• [Presentation] Bioimaging devices for cell transplantation therapy in regenerative medicine (2016)
  • Author(s): D. Onoshima
  • Organizer: 8th International Symposium on Advanced Plasma Science and its Applications for Nitrides and Nanomaterials (ISPlasma2016)
  • Place of Presentation: Nagoya University, Aichi, Japan
  • Year and Date: 2016-03-06
  • Int'l Joint Research / Invited
• [Presentation] Single cancer cell isolation and visualization in microfluidic chip for cytoscreening of cancer stem cells (2015)
  • Author(s): D. Onoshima, H. Yukawa, and Y. Baba
  • Organizer: World Molecular Imaging Congress (WMIC) 2015
  • Place of Presentation: Hawaii Convention Center, Honolulu, Hawaii
  • Year and Date: 2015-09-01
  • Int'l Joint Research
• [Presentation] Nanobiodevices for diagnostics, therapeutics, and future mobility (2015)
  • Author(s): D. Onoshima
  • Organizer: 6th International Conference on Nanotechnology: Fundamentals and Applications (ICNFA’15)
  • Place of Presentation: Alimara Hotel, Barcelona, Spain
  • Year and Date: 2015-07-15
  • Int'l Joint Research / Invited
• [Presentation] モビリティと融合したナノバイオ計測技術開発への取り組み (2015)
  • Author(s): 小野島大介
  • Organizer: MERRO「第1回次世代若手－医・理・工連携－研究会」in蒲郡, 夢の医療技術構想（グランドチャレンジ）再生医療と再生治療・がんの微小環境・地域産業との連携
  • Place of Presentation: 蒲郡市民会館
  • Year and Date: 2015-01-17 – 2015-01-18
  • Invited
• [Presentation] Microfluidic Stretching of DNA with fluorescent gold nanoparticle for optical/electron microscopic imaging of a single DNA methylation (2014)
  • Author(s): D. Takeshita, D. Onoshima, H. Yukawa, T. Yasui, N. Kaji, and Y. Baba
  • Organizer: The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2014)
  • Place of Presentation: Henry B. Gonzalez Convention Center, San Antonio, USA
  • Year and Date: 2014-10-26 – 2014-10-30
• [Presentation] Sizing and sorting of single DNA molecules by microfluidic molecular combing device (2014)
  • Author(s): D. Onoshima, N. Kawakita, D. Takeshita, H. Yukawa, and Y. Baba
  • Organizer: The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2014)
  • Place of Presentation: Henry B. Gonzalez Convention Center, San Antonio, USA
  • Year and Date: 2014-10-26 – 2014-10-30
• [Presentation] Trapping, stretching, and separating single DNA molecules on a chip (2014)
  • Author(s): D. Onoshima
  • Organizer: EMN Summer Meeting 2014
  • Place of Presentation: The Westin Resort & Spa, Cancun, Mexico
  • Year and Date: 2014-06-09 – 2014-06-12
  • Invited
• [Presentation] 金ナノ粒子-蛍光分子複合体を用いたメチル化DNAの１分子伸長・画像解析デバイス (2014)
  • Author(s): 竹下大貴、小野島大介、湯川博、安井隆雄、加地範匡、馬場嘉信
  • Organizer: 化学とマイクロ・ナノシステム学会第29回研究会
  • Place of Presentation: 日本女子大学 目白キャンパス
  • Year and Date: 2014-05-22 – 2014-05-23
• [Book] 医療・診断・創薬の化学 (2017)
  • Author(s): 日本化学会 編
  • Total Pages: 208
  • Publisher: 化学同人
  • ISBN: 9784759813845
• [Patent(Industrial Property Rights)] 細胞捕捉フィルター (2016)
  • Inventor(s): 小野島大介、他9名
  • Industrial Property Rights Holder: 旭硝子株式会社、国立大学法人名古屋大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-09-23
  • Overseas
• [Patent(Industrial Property Rights)] 細胞外小胞の回収方法 (2016)
  • Inventor(s): 馬場嘉信、小野島大介、他5名
  • Industrial Property Rights Holder: 旭硝子株式会社、国立大学法人名古屋大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2016-046011
  • Filing Date: 2016-03-09
• [Patent(Industrial Property Rights)] 細胞捕捉チップ及び細胞捕捉方法 (2015)
  • Inventor(s): 龍腰健太郎、小野島大介、他7名
  • Industrial Property Rights Holder: 旭硝子株式会社、国立大学法人名古屋大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2015-045829
  • Filing Date: 2015-03-09