High-throughput droplet sorting devices for large-scale single-cell analysis

• Project/Area Number: 18K14101
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 28050:Nano/micro-systems-related
• Research Institution: The University of Tokyo
• Principal Investigator: Isozaki Akihiro 東京大学, 大学院理学系研究科(理学部), 特任助教 (10732555)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2019: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
• Keywords: Droplet microfluidics / Single-cell analysis / マイクロ流路 / Flow cytometry / 液滴 / 抗体産生 / 創薬

Outline of Final Research Achievements

I have developed a high-speed large droplet manipulation method that is an order of magnitude faster than conventional methods. Specifically, by precisely controlling the timing and position of the force applied to droplets, the developed method enables gentle and fast manipulation of fragile, large droplets, resulting in a 20-fold increase in the manipulation speed compared to conventional methods. Furthermore, I have demonstrated that microalgal cells can move freely in droplets larger than 100 pL, which is an advantage of our method. The survival rate of their microalgal cells and leukemia cells is increased as well as the antibody production of hybridoma cells. Furthermore, I have used it to isolate microorganisms with a slow growth rate, which only exist in about 2% of the population, confirming its usefulness and versatility.

Academic Significance and Societal Importance of the Research Achievements

液滴分取スピードの高速化により、これまで見つけられなかったような希少な細胞を発見・単離できる可能性が大幅に向上する。これにより、微生物学、免疫学、遺伝学などの基礎科学における新たな発見や、精密医療、ウイルス検査、創薬、バイオ燃料開発、スマートセル産業などにおける効率化技術の開発といった、さまざまな応用展開が期待される。

Research Products

• [Int'l Joint Research] LMU München(ドイツ)
• [Journal Article] Dual sequentially addressable dielectrophoretic array for high-throughput, scalable, multiplexed droplet sorting (2021)
  • Author(s): Isozaki Akihiro、Huang Dunhou、Nakagawa Yuta、Goda Keisuke
  • Journal Title: Microfluidics and Nanofluidics Volume: 25 Issue: 4 Pages: 1-10
  • DOI: 10.1007/s10404-021-02432-z
  • Peer Reviewed / Open Access
• [Journal Article] Sequentially addressable dielectrophoretic array for high-throughput sorting of large-volume biological compartments (2020)
  • Author(s): A Isozaki, Y Nakagawa, M H Loo, Y Shibata, N Tanaka, D L Setyaningrum, J-W Park, Y Shirasaki, H Mikami, D Huang, H Tsoi, C T Riche, T Ota, H Miwa, Y Kanda, T Ito, K Yamada, O Iwata, K Suzuki, S Ohnuki, Y Ohya, Y Kato, T Hasunuma, S Matsusaka, M Yamagishi, M Yazawa, S Uemura, K Nagasawa, H Watarai, D Di Carlo, K Goda
  • Journal Title: Science Advances Volume: 29 Issue: 22 Pages: 1-11
  • DOI: 10.1126/sciadv.aba6712
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] "High-throuhgput sorting of nanoliter droplets using an electrode array with a slanted microchannel (2020)
  • Author(s): Mun Hong Loo, Yuta Nakagawa, Akihiro Isozaki, and Keisuke Goda
  • Organizer: The 24th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS2020)
  • Int'l Joint Research
• [Presentation] Sequentially addressable dielectrophoretic array for high-throughput large-droplet sorting (2020)
  • Author(s): Akihiro Isozaki, Yuta Nakagawa, Mun Hong Loo, Dino Di Carlo, and Keisuke Goda
  • Organizer: CYTO Virtual 2020
  • Int'l Joint Research
• [Presentation] Exploring the advantage of large droplets for single-cell analysis (2020)
  • Author(s): Yuta Nakagawa, Shinsuke Ohnuki, Akihiro Isozaki, Naoko Kondo, Yoshikazu Ohya, and Keisuke Goda
  • Organizer: 化学とマイクロ・ナノシステム学会第42回研究会
• [Presentation] 電極アレイを用いた高速液滴分取 (2019)
  • Author(s): 磯崎 瑛宏、中川 悠太、Mun Hong Loo、芝田 悠大、田中 直樹、Dwi Larasati Setyaningrum、Jee-Woong Park、白崎 善隆、三上 秀治、上村 想太郎、Dino Di Carlo、合田圭介
  • Organizer: 化学とマイクロ・ナノシステム学会
• [Presentation] ow-loss droplet picking for high-throughput rare-cell sorting (2019)
  • Author(s): Chenqi Zhang, Mun Houng Loo, Yuta Nakagawa, Akihiro Isozaki, Keisuke Goda
  • Organizer: 化学とマイクロ・ナノシステム学会
• [Presentation] A droplet microfluidic platform for separation of cells with low proliferative activity (2019)
  • Author(s): Yuta Nakagawa, Mun Hong Loo, Shinsuke Ohnuki, Yoshikazu Ohya, Akihiro Isozaki, Keisuke Goda
  • Organizer: 化学とマイクロ・ナノシステム学会
• [Presentation] 電極アレイを用いた多分岐高速液滴分取 (2019)
  • Author(s): 磯崎 瑛宏, Dunhou Huang, 合田 圭介
  • Organizer: 化学とマイクロ・ナノシステム学会
• [Presentation] 電極アレイを用いた超高速液滴分取とその応用 (2019)
  • Author(s): 中川 悠太
  • Organizer: 日本化学会春季年会
• [Presentation] An electrode-array-based high-throughput nano-liter-droplet sorter assisted by a slanted microchannel (2019)
  • Author(s): Loo Mun Hong
  • Organizer: 日本化学会春季年会
• [Presentation] 微小液滴内におけるハイブリドーマの抗体産生量 (2018)
  • Author(s): 磯崎瑛宏
  • Organizer: 化学とマイクロ・ナノシステム学会第38回研究会
• [Remarks] Akihiro Isozaki
  • URL: https://sites.google.com/site/akihiroisozaki/