3D molecular orientation imaging for in vivo visualizations of denatured proteins

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K19346
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 47:Pharmaceutical sciences and related fields
• Research Institution: Juntendo University
• Principal Investigator: Otomo Kohei 順天堂大学, 大学院医学研究科, 准教授 (40547204)
• Project Period (FY): 2021-07-09 – 2024-03-31
• Keywords: 二光子顕微鏡法 / 超解像顕微鏡法 / 分子配向イメージング / 高次構造 / 偏光イメージング / 一分子イメージング

Outline of Final Research Achievements

The objective of this research project is to establish a novel three-dimensional molecular orientation imaging method to realize simultaneous visualization of localization and higher-order structure of biomolecules. To achieve it, a novel two-photon microscopy system based on polarization anisotropy analysis and super-resolution microscopy techniques was developed. Proof-of-principle experiments using optical phantoms such as fluorescent beads and fluorophore-labeled liposomes, have established the technical basis for higher-order structural imaging of biomolecules. Experiments using living cells are currently in progress and results will be reported in due course. In the course of our research, we have also succeeded in developing auxiliary elemental technologies, some of which were reported as the original papers.

Free Research Field

物理薬学

Academic Significance and Societal Importance of the Research Achievements

生体は分子、細胞小器官、細胞、組織の多階層において精微な三次元構造を持ち、この多階層における様々な『かたち』は機能と密接な関わりを持つ。これを理解するイメージング技術の発展は目覚ましく、各階層における現象の詳細記述が可能になりつつある。一方で、個々の方法論には、原理に応じた限界が存在し、階層間の相互理解の妨げとなっている。本研究課題は組織から細胞小器官までを主対象とする蛍光顕微鏡法にて、分子のかたちの可視化する基盤技術の開発に挑んだ。本技術成熟の暁には、生体を構成する多階層構造の縦断的理解が可能となり、生命科学研究における多大なインパクトが期待される。