Development of analyzer for physiological response of biological tissue under localized stress by near-field fluorescence microscopy

2015 Fiscal Year Final Research Report

• Project/Area Number: 25286029
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Nanobioscience
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: Norihiro Umeda 東京農工大学, 工学(系)研究科(研究院), 教授 (60111803)
• Co-Investigator(Kenkyū-buntansha): OHTA Yoshihiro 東京農工大学, 大学院工学研究院, 准教授 (10223843) ; IWAMI Kentaro 東京農工大学, 大学院工学研究院, 准教授 (80514710)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: ミトコンドリア / ベント型光ファイバー / 局所微小荷重 / 蛍光色素 / 共焦点顕微鏡 / ヤング率 / ナノニュートン / ROS

Outline of Final Research Achievements

Mitochondria produce reactive oxygen spices (ROS). Especially, in the case of ischemia- reperfusion injury, physical stress induced by blood flow at narrowing blood vessel causes ROS production from mitochondria in the living cell. However, the relationship between physical stress applied to mitochondrion and mitochondrial ROS production is absolutely unknown. To investigate this relationship, we developed a force-applying confocal laser scanning microscopy (F-CLSM). The ROS response of mitochondrion was measured by using MitoSOX fluorescent indicator. We confirmed 90 % difference of fluorescence increase rates between inhibitor and activator injections. Finally, we monitored the fluorescence of MitoSOX while applying force onto single mitochondrion. In the case of applied force of 40 nN, fluorescence intensity rate was larger than that of 10 nN. Therefore, we revealed that the ROS produced in the mitochondrion depends on the magnitude of physical stress applied to mitochondrion.

Free Research Field

ナノフォトニクス