Efficient targeting method for correlative light and electron microscopy

2023 Fiscal Year Final Research Report

• Project/Area Number: 22K20741
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0802:Biomedical structure and function and related fields
• Research Institution: Yamaguchi University
• Principal Investigator: Yoshimura Azumi 山口大学, 大学院医学系研究科, 講師 (90467340)
• Project Period (FY): 2022-08-31 – 2024-03-31
• Keywords: 電子顕微鏡 / 光-電子相関顕微鏡法

Outline of Final Research Achievements

Correlative light and electron microscopy (CLEM) enables correlation of molecular and structural information and allows qualitative and quantitative analysis of regions of interest (ROI) specified by light microscopy. The correlation however becomes challenging as the specimen size increases, due to the difficulty in ROI targeting after EM sample preparation. This study aims to develop an efficient ROI targeting method for CLEM that can be performed with widely used fluorescence microscopy. Using a fluorescent label detectable both before and after EM samples preparation, landmark structures were introduced. Based on their positions detected by fluorescence microscopy at different steps of the sample preparation, step-wise ROI targeting was achieved.

Free Research Field

光-電子相関顕微鏡法

Academic Significance and Societal Importance of the Research Achievements

立体的に大きな試料における光-電子相関顕微鏡法（CLEM）には、目的領域のターゲティングに先端的設備を要する、電子観察のデータサイズが増大しスループットが低くなるなどの問題点がある。本課題で実現されたターゲティング法は、一般的な蛍光観察のみを用いて実行可能なもので、手法の簡略化、自動化を図り汎用性を高めることで、3次元でのCLEMを効率化することが期待される。