Development of a method for the epigenetic analysis of single cells under a micro fluidic device

• Project/Area Number: 25286035
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Nano/Microsystems
• Research Institution: The University of Tokyo
• Principal Investigator: Oana Hidehiro 東京大学, 工学(系)研究科(研究院), 准教授 (20314172)
• Co-Investigator(Renkei-kenkyūsha): WASHIZU MASAO 東京大学, 大学院工学系研究科, 教授 (10201162) ; OKEYO KENNEDY 東京大学, 大学院工学系研究科, 助教 (10634652)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥19,760,000 (Direct Cost: ¥15,200,000、Indirect Cost: ¥4,560,000); Fiscal Year 2015: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2014: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2013: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000)
• Keywords: ゲノム / DNA / マイクロ流体デバイス / バイオテクノロジー / マイクロ・ナノデバイス / シングルセル / エピジェネティクス

Outline of Final Research Achievements

We developed a new methodology for analysis of individual intact chromosomes/chromatins isolated from single cells. We developed a custom-designed microfluidic device for isolation, control of the higher order structure of individual chromosome/chromatin by changing solution conditions under a fluorescence microscope. This device is also able to perform immunofluorescence staining. The method developed here has a potential application for direct investigation of the epigenetic profile of intact chromatin fibers at individual cell level.

Research Products

• [Journal Article] Non-destructive handling of individual chromatin fibers isolated from single cells in a microfluidic device utilizing an optically driven microtool. (2013)
  • Author(s): Oana H, Nishikawa K, Matsuhara H, Yamamoto A, Yamamoto TG, Haraguchi T, Hiraoka Y, Washizu M.
  • Journal Title: Lab on a Chip. Volume: 14 Issue: 4 Pages: 696-704
  • DOI: 10.1039/c3lc51111a
  • Peer Reviewed
• [Presentation] A MICROFLUIDIC PLATFORM FOR ANALYSIS OF SINGLE CHROMATIN FIBERS: TOWARD SINGLE CELL EPIGENETIC STUDY (2016)
  • Author(s): Hidehiro Oana, Hiroki Mori, Kennedy O. Okeyo and Masao Washizu
  • Organizer: The Eighth International Conference on Microtechnologies in Medicine and Biology
  • Place of Presentation: 大韓民国, ソウル
  • Year and Date: 2016-04-20
  • Int'l Joint Research
• [Presentation] Development of a Method for Investigation of Individual Intact Chromatin Fibers Isolated from Single Cells in a Microfluidic Channel (2016)
  • Author(s): Hidehiro Oana, Hiroki Mori, Kennedy O. Okeyo and Masao Washizu
  • Organizer: SELECTBIO Conference, Single Molecule & Single Cell Analysis
  • Place of Presentation: スペイン王国, マドリッド
  • Year and Date: 2016-03-15
  • Int'l Joint Research
• [Presentation] マイクロ流路を用いたクロマチン展開挙動の計測法の開発 (2016)
  • Author(s): 森 裕貴，オケヨ ケネディ オモンディ， 鷲津 正夫，小穴 英廣
  • Organizer: 日本機械学会第28回バイオエンジニアリング講演会
  • Place of Presentation: 東京工業大学大岡山キャンパス 東京都目黒区
  • Year and Date: 2016-01-10
• [Presentation] Non-destructive Handling of Individual Intact Chromatin Fibers Isolated from Single Cells in a Microfluidic Device - Toward Single Cell Epigenetic Study by Microscopy (2015)
  • Author(s): Hidehiro Oana
  • Organizer: 15th Asia-Pacific International Symposium on Microscale Separations and Analysis
  • Place of Presentation: National Cheng Kung University, Tainan, Taiwan
  • Year and Date: 2015-11-15
  • Int'l Joint Research / Invited
• [Presentation] Immunofluorescence Staining and Extending of Individual Intact Chromatin Fibers Isolated from Single Cells utilizing a Microfluidic Device (2014)
  • Author(s): Hidehiro Oana
  • Organizer: The 8th International Workshop on Approaches to Single-Cell Analysis
  • Place of Presentation: ケニア，ナイロビ市
  • Year and Date: 2014-09-11
  • Invited
• [Presentation] マイクロ流体デバイスを用いた動物細胞のクロマチンファイバー単離・伸長手法の研究 (2014)
  • Author(s): 伊藤哲久・ケネディ オケヨ ・鷲津正夫 ・小穴英廣
  • Organizer: 第29回化学とマイクロ・ナノシステム学会
  • Place of Presentation: 東京都文京区日本女子大学
  • Year and Date: 2014-05-23
• [Presentation] 相同組換えタンパクをプローブに用いた標的塩基配列の顕微鏡下リアルタイム探索 (2014)
  • Author(s): 東郷智之 ・ケネディ オケヨ ・小穴英廣 ・鷲津正夫
  • Organizer: 第29回化学とマイクロ・ナノシステム学会
  • Place of Presentation: 東京都文京区日本女子大学
  • Year and Date: 2014-05-23
• [Presentation] Development of an Optically Driven Microtool for Micromanipulation and Investigation of Individual Chromatin Fibers in a Microfluidic Device (2014)
  • Author(s): Hidehiro Oana
  • Organizer: Advances in Microfluidics & Nanofluidics 2014 (AMN2014)
  • Place of Presentation: 中華民国，台北市
  • Year and Date: 2014-05-21
  • Invited
• [Presentation] 抗体修飾マイクロビーズを用いたゲノムDNA・蛋白質複合体の単分子操作技術の開発
  • Author(s): 若林貴之ら
  • Organizer: 第27回化学とマイクロ・ナノシステム研究会
  • Place of Presentation: 宮城県仙台市東北大学
• [Presentation] 微細操作技術のシングルセル・エピゲノム解析への応用
  • Author(s): 小穴英廣
  • Organizer: 第2回イノバイオ研究会セミナー
  • Place of Presentation: 東京都文京区東京大学
  • Invited
• [Presentation] 微細加工・微細操作技術のシングルセルベース ゲノム研究への応用
  • Author(s): 小穴英廣
  • Organizer: 次世代マイクロ化学チップ研究会
  • Place of Presentation: 東京都文京区東京大学
  • Invited
• [Presentation] NON-INVASIVE HANDLING OF CHROMATIN FIBERS ISOLATED FROM INDIVIDUAL CELLS IN A MICROCHANNEL UTILIZING AN OPTICALLY DRIVEN MICROTOOL
  • Author(s): 小穴英廣ら
  • Organizer: 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences
  • Place of Presentation: ドイツ連邦共和国フライブルク