Super-resolution oxygen partial pressure imaging of single cells

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K12762
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 90110:Biomedical engineering-related
• Research Institution: Keio University
• Principal Investigator: Tsukada Kosuke 慶應義塾大学, 理工学部(矢上), 教授 (00351883)
• Co-Investigator(Kenkyū-buntansha): 毛利 聡 川崎医科大学, 医学部, 教授 (00294413)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: 超解像 / 酸素分圧 / イメージング / 代謝

Outline of Final Research Achievements

The establishment of a method to visualize oxygen concentration information in living cells with spatial resolution beyond the diffraction limit will contribute to the elucidation of local life phenomena. A laser beam was split to generate a Gaussian beam and a doughnut-shaped beam. The object was scanned two-dimensionally with each beam, and super-resolution images were obtained by subtracting the obtained images. Optical conditions were determined to minimize the loss of intensity information during the differencing process and to obtain high spatial resolution. As a result, a resolution of approximately 100 nm was achieved based on experimental measurements. Furthermore, an algorithm to rapidly determine the phosphorescence lifetime for quantifying the oxygen partial pressure enables fast imaging. Taken together, the proposed method is expected to be used for fast super-resolution oxygen partial pressure imaging of living cells.

Free Research Field

生体医工学

Academic Significance and Societal Importance of the Research Achievements

肺で取り込まれた分子状酸素は血管から抹消組織に拡散によって移動し，細胞内のミトコンドリアで消費されてエネルギー産生に利用される．その細胞内の局所空間における酸素濃度情報がどのように生理機能や病態に関わるのか大変興味深いが，測定技術の欠落から未解明のままである．この研究では細胞内の酸素濃度情報を光学的に取得するための方法を提案し，数値シミュレーションと実測実験から有効性を実証した．