Single-molecule fluorescence detection of kinesin and ATP using nanotracks

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K13644
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Nanobioscience
• Research Institution: Kyoto University
• Principal Investigator: Yokokawa Ryuji 京都大学, 工学研究科, 准教授 (10411216)
• Co-Investigator(Renkei-kenkyūsha): IINO Ryota 分子科学研究所, 生命・錯体分子科学研究領域 生体分子機能研究部門, 教授 (70403003) ; TOMISHIGE Michio 青山学院大学, 理工学部, 教授 (50361530)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: マイクロ・ナノデバイス / 分子モーター / 生物物理 / ナノバイオ / 1分子計測（SMD）

Outline of Final Research Achievements

We developed Linear Zero Mode Waveguides (LZMWs) for simultaneous observation of kinesin and ATP under higher concentration of fluorescently labeled ATP than that used in the conventional total internal reflection microscopy (TIRFM). Single-molecule fluorescent microscopy is a promising method to analyze correlation between the mechanical displacement and hydrolysis of ATP by a kinesin molecule. However, the concentration of fluorescently labeled ATP was limited up to 10 nM in TIRFM, thus available data from experiment was limited. Proposed method enables to use ten times higher concentration of fluorescently labeled ATP by confining excitation lights in LZMWs, which are nano-slits with 100 nm in width.

Free Research Field

BioMEMS、MicroTAS