High-speed single cell diagnosis by atomic force microscopy

• Project/Area Number: 21H01787
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 29010:Applied physical properties-related
• Research Institution: Hokkaido University
• Principal Investigator: Okajima Takaharu 北海道大学, 情報科学研究院, 教授 (70280998)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2023: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2022: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2021: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000)
• Keywords: 原子間力顕微鏡 / 細胞 / 原子間力顕微 / 細胞診断 / メカニクス / 細胞力学診断

Outline of Research at the Start

原子間力顕微鏡（AFM）は、生細胞・組織の力学特性を高い時間空間分解能で計測することができる。本研究では、AFMを用いて接着状態の細胞の疾患を1細胞単位で定量、且つ高速に同定する新手法を開発する。令和3年度は、細胞レオロジー計測の高速化技術、およびボルツマンの重畳原理に基づく細胞レオロジー解析法の基礎技術および概念を利用し、高速な１細胞レオロジー診断システムを開発する。令和4年度は、細胞株を用いて１細胞単位の高速診断技術を開発する。令和5年度は、プライマリー細胞・組織を用いて高速な細胞疾患の診断技術を開発する。

Outline of Final Research Achievements

Atomic force microscopy (AFM)-methods have been studied to quantify and rapidly identify the progression of adherent cancer cells at the single-cell level. The high-speed single-cell rheology diagnosis technique has been developed by using AFM-cell rheology analysis and the Boltzmann's superposition principle. We also investigated application to disease cells other than cancer cells and multicellular population systems.

Academic Significance and Societal Importance of the Research Achievements

がん細胞を1細胞単位で診断する技術は、究極のがん早期発見技術と言っても過言ではない。近年、がん細胞の進行度と力学特性との関係性が指摘されている。本研究では、原子間力顕微鏡（AFM）を用いて、接着状態のがん細胞の進行度を1細胞単位で定量、且つ高速に同定する新手法を開発した。細胞レオロジー計測の高速化技術とボルツマンの重畳原理に基づく細胞レオロジー解析法の基礎技術および概念を利用し、高速な１細胞レオロジー診断技術を開発した。また、がん細胞以外の疾患細胞や多細胞集団系への適用を検討した。

Research Products

• [Journal Article] Compressed sensing reconstruction of cell mechanical images obtained from atomic force microscopy (2024)
  • Author(s): Yuki Miyata, Feng-Yueh Chan, Takayuki Uchihashi, Takaharu Okajima
  • Journal Title: Japanese Journal of Applied Physics Volume: - Issue: 4 Pages: 04SP46-04SP46
  • DOI: 10.35848/1347-4065/ad34de
  • Peer Reviewed
• [Journal Article] Local intracellular stiffening of ascidian embryo in cleavage developmental stage observed by atomic force microscopy (2024)
  • Author(s): Takahiro Kotani, Yuki Miyata, Yosuke Tsuboyama, Yuki Fujii, Takaharu Okajima
  • Journal Title: Japanese Journal of Applied Physics Volume: - Issue: 4 Pages: 04SP64-04SP64
  • DOI: 10.35848/1347-4065/ad3760
  • Peer Reviewed
• [Journal Article] Atomic force microscopy for investigating cell and tissue mechanics as heterogeneous and hierarchical materials (2023)
  • Author(s): Takaharu OKAJIMA, Kaori KURIBAYASHI-SHIGETOMI
  • Journal Title: Journal of Biomechanical Science and Engineering Volume: 18 Issue: 4 Pages: 23-00339-23-00339
  • DOI: 10.1299/jbse.23-00339
  • ISSN: 1880-9863
  • Peer Reviewed / Open Access
• [Journal Article] Pathogenic Roles of Cardiac Fibroblasts in Pediatric Dilated Cardiomyopathy (2023)
  • Author(s): Tsuru Hirofumi、Yoshihara Chika、Suginobe Hidehiro、Matsumoto Mizuki、Ishii Yoichiro、Narita Jun、Ishii Ryo、Wang Renjie、Ueyama Atsuko、Ueda Kazutoshi、Hirose Masaki、Hashimoto Kazuhisa、Nagano Hiroki、Tanaka Ryosuke、Okajima Takaharu、Ozono Keiichi、Ishida Hidekazu
  • Journal Title: Journal of the American Heart Association Volume: 12 Issue: 13
  • DOI: 10.1161/jaha.123.029676
  • Peer Reviewed / Open Access
• [Journal Article] Mechanical properties of epithelial cells in domes investigated using atomic force microscopy (2023)
  • Author(s): Kenta Shigemura, Kaori Kuribayashi-Shigetomi, Ryosuke Tanaka, Haruka Yamasaki, and Takaharu Okajima
  • Journal Title: Frontiers in Cell and Developmental Biology Volume: 11 Pages: 1245296-1245296
  • DOI: 10.3389/fcell.2023.1245296
  • Peer Reviewed / Open Access
• [Journal Article] Atomic force microscopy identifies the alteration of rheological properties of the cardiac fibroblasts in idiopathic restrictive cardiomyopathy (2022)
  • Author(s): Mizuki Matsumoto, Hirofumi Tsuru, Hidehiro Suginobe, Jun Narita, Ryo Ishii, Masaki Hirose, Kazuhisa Hashimoto, Renjie Wang, Chika Yoshihara, Atsuko Ueyama, Ryosuke Tanaka, Keiichi Ozono, Takaharu Okajima, Hidekazu Ishida
  • Journal Title: PLoS ONE Volume: 17 Issue: 9 Pages: e0275296-e0275296
  • DOI: 10.1371/journal.pone.0275296
  • Peer Reviewed / Open Access
• [Journal Article] Atomic force microscopy identifies cellular elasticity of pulmonary arterial smooth muscle cells in a patient with idiopathic pulmonary arterial hypertension (2021)
  • Author(s): Shinichi Katsuragi, Nao Tatsumi, Mizuki Matsumoto, Jun Narita, Ryo Ishii, Hidehiro Suginobe, Hirofumi Tsuru, Wang Renjie, Shigetoyo Kogaki, Keiichi Ozono, Ryosuke Tanaka, Takaharu Okajima, and Hidekazu Ishida
  • Journal Title: Cardiology Research Volume: 12 Issue: 4 Pages: 231-237
  • DOI: 10.14740/cr1282
  • Peer Reviewed / Open Access
• [Presentation] 大腸がん微小環境における線維芽細胞の物理測定 (2024)
  • Author(s): 山﨑遼、繁富（栗林）香織、岡嶋孝治
  • Organizer: 日本顕微鏡学会・SPM研究会2023
• [Presentation] Rheological properties of Submucosal and subperitoneal fibroblasts measured by atomic force microscopy (2023)
  • Author(s): H.Yamasaki, K.Kuribayashi-Shigetomi, M.Kojima, T.Okajima
  • Organizer: 第61回日本生物物理学会年会
• [Presentation] Atomic force microscopy reveals a relationship between rheological properties of cells and cell fates in the early developmental stages (2023)
  • Author(s): Y. Miyata, T. Kotani, Y. Tsuboyama, T. Matsuo, T. Okajima
  • Organizer: 31st International Colloquium on Scanning Probe Microscopy
  • Int'l Joint Research
• [Presentation] Spatiotemporal dynamics of single-cell rheology in developing embryos observed by atomic force microscopy (2023)
  • Author(s): T. Kotani, Y. Miyata, Y. Tsuboyama, T. Matsuo, M. Yokobori, Y. Fujii, T. Okajima
  • Organizer: 31st International Colloquium on Scanning Probe Microscopy
  • Int'l Joint Research
• [Presentation] Embryo mechanics probed by atomic force microscopy (2023)
  • Author(s): T. Okajima
  • Organizer: 第61回日本生物物理学会年会
  • Invited
• [Presentation] 多細胞メカニクスの時空間マッピング測定 (2022)
  • Author(s): 岡嶋 孝治
  • Organizer: ナノプローブテクノロジー第167委員会第103回研究会
  • Invited
• [Presentation] 単層構造・3D構造を有する上皮細胞の走電性の観察 (2022)
  • Author(s): 小野実央, 小川美優, 永野真尭, 岡嶋孝治
  • Organizer: 第83回応用物理学会秋季学術講演会
• [Presentation] パルスフォース原子間力顕微鏡による上皮細胞シートの 力学特性の測定 (2022)
  • Author(s): 垣内琢規, 土生飛翔, 岡嶋孝治
  • Organizer: 第83回応用物理学会秋季学術講演会
• [Presentation] Quantifying tension and elastic modulus of epithelial cells in 3D dome by atomic force microscopy (2022)
  • Author(s): K. Shigemura, K. Kuribayashi-Shigetomi, A. Subagyo, K. Sueoka and T. Okajima
  • Organizer: The 22nd International Vacuum Congress (IVC-22)
  • Int'l Joint Research
• [Presentation] Simulating stress relaxation curves of soft materials obtained in atomic force microscopy experiments (2022)
  • Author(s): M. Nakada, Y. Tsuboyama, and T. Okajima
  • Organizer: The 22nd International Vacuum Congress (IVC-22)
  • Int'l Joint Research
• [Presentation] Nanomechanics of single cells and tissues revealed by atomic force microscopy (2021)
  • Author(s): T. Okajima
  • Organizer: Pacifichem 2021, Experimental and Computational Analysis of the Nano-Bio Interface for Sustainable Nanotechnology
  • Int'l Joint Research / Invited
• [Presentation] Quantifying mechanical property of epithelial cell dome by atomic force microscopy (2021)
  • Author(s): K. Shigemura*, K.K. Shigetomi, A. Subagyo, K. Sueoka and T. Okajima,
  • Organizer: 29th International Colloquium on Scanning Probe Microscopy (ICSPM29)
• [Presentation] A numerical simulation of stress relaxation atomic force microscopy (2021)
  • Author(s): M. Nakada* and T. Okajima,
  • Organizer: 29th International Colloquium on Scanning Probe Microscopy (ICSPM29)
• [Presentation] 原子間力顕微鏡による上皮ドームの力学解析 (2021)
  • Author(s): 茂村研太、繁富（栗林）香織、スバギョ・アグス、末岡和久、岡嶋孝治
  • Organizer: 第59回日本生物物理学会年会
• [Presentation] Observations of structural change in epithelial dome under constant electric field (2021)
  • Author(s): Mataka Nagano, Miyu Ogawa, Yuki Fujii, Takaharu Okajima
  • Organizer: 第59回日本生物物理学会年会