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Nanometer scale resolution imaging of pH and reactive oxygen species on the outer membrane surface for fast and effective cell discrimination in cancer tissues.

Research Project

Project/Area Number 19K05228
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 28040:Nanobioscience-related
Research InstitutionKanazawa University

Principal Investigator

Puppulin Leonardo  金沢大学, ナノ生命科学研究所, 特任助教 (40771580)

Co-Investigator(Kenkyū-buntansha) 山田 忠明  京都府立医科大学, 医学(系)研究科(研究院), 准教授 (00507048)
細木 誠之  京都薬科大学, 薬学部, 准教授 (30433254)
Project Period (FY) 2019-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
KeywordsRaman spectroscopy / Nanosensors / pH / Hydrogen peroxide / cell surface / cell surface pH / nanosensor / SERS / hydrogen peroxides / nano-sensor / redox biology / Reactive Oxygen Species / Imaging / Cancer cell
Outline of Research at the Start

We will develop a nanometer scale sensor of pH and reactive oxygen species to be anchored to the outer membrane surface of cells for fast and effective discrimination of cancer cell type in tissues.

Outline of Final Research Achievements

We developed and tested a nano-sensor for quantification of hydrogen peroxide (H2O2) on the cell surface. The nanosensor is based on bioconjugated gold nanoparticles that can be anchored to the extracellular cell surface and can quantify H2O2 using surface enhanced Raman spectroscopy (SERS). We successfully tested the new nanosensor on adenocarcinomic human alveolar basal epithelial cells (A59 cell line) for highly localized quantitative measurements of H2O2. In addition, we optimized the use of our previously developed pH nanosensor for cell surface pH imaging. Using a laser line illumination mode, we simultaneously acquired 400 SERS spectra from a wide area of the cell sample. We analyzed different cell lines: pancreatic cancer cell BxPC; pancreatic cancer cells AsPC-1; murine fibroblast cells NIH/3T3; human fibroblast WI-38; lung cancer A549.

Academic Significance and Societal Importance of the Research Achievements

Quantification of pH and hydrogen peroxide on the cell surface is critically important to study many cellular functions and can be exploited to discriminate cancer cells from normal cells in tissues. The nanosensors developed in our research project showed great potential to accomplish these goals.

Report

(4 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • Research Products

    (2 results)

All 2021 2020

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Presentation (1 results) (of which Invited: 1 results)

  • [Journal Article] Plasma membrane anchored nanosensor for quantifying endogenous production of H2O2 in living cells2021

    • Author(s)
      Hosogi Shigekuni、Marunaka Yoshinori、Ashihara Eishi、Yamada Tadaaki、Sumino Ayumi、Tanaka Hideo、Puppulin Leonardo
    • Journal Title

      Biosensors and Bioelectronics

      Volume: 179 Pages: 113077-113077

    • DOI

      10.1016/j.bios.2021.113077

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Presentation] Surface enhanced Raman spectroscopy probe for nanometer-scale measurement of pH and hydrogen peroxides on the outer membrane of cells2020

    • Author(s)
      Puppulin Leonardo
    • Organizer
      The 97th Annual Meeting of the Physiological Society of Japan
    • Related Report
      2019 Research-status Report
    • Invited

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Published: 2019-04-18   Modified: 2023-01-30  

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