A Nanodiamond-peptide Bioconjugate for Fluorescence and ODMR Microscopy of a Single Actin Filament
A Nanodiamond-peptide Bioconjugate for Fluorescence and ODMR Microscopy of a Single Actin Filament
复制标题
DOI：
10.2116/analsci.32.1165
复制
发表时间：
2016-11-01
影响因子：
1.6
通讯作者：
Shirakawa, Masahiro
中科院分区：
文献类型：
--
作者：
Genjo, Takuya;Sotoma, Shingo;Shirakawa, Masahiro
近年来，机械刺激引起肌动蛋白细丝构象变化的重要性日益被人们所认识，特别是在机械生物学方面。然而，尽管单个肌动蛋白细丝具有基本的重要性，但由于传统荧光分子的低光稳定性，用荧光显微镜长期观察它一直是困难的。本文报道了一种新型的肌动蛋白细丝分子标记系统，该系统使用带有氮空位中心的荧光纳米金刚石(ND)颗粒；ND具有灵活的化学可修饰性、极高的光稳定性和生物相容性，并通过光学检测磁共振(ODMR)测量定量地提供各种物理信息。我们用肌动蛋白细丝特异性结合肽Lifeact对ND进行了化学表面修饰，并通过ODMR选择性成像方法观察到纯Lifeact修饰的ND和肌动蛋白细丝的共存，表明利用ND颗粒可以长期观察和定量分析单个分子。
Recently, the importance of conformational changes in actin filaments induced by mechanical stimulation of a cell has been increasingly recognized, especially in terms of mechanobiology. Despite its fundamental importance, however, long-term observation of a single actin filament by fluorescent microscopy has been difficult because of the low photostability of traditional fluorescent molecules. This paper reports a novel molecular labeling system for actin filaments using fluorescent nanodiamond (ND) particles harboring nitrogen-vacancy centers; ND has flexible chemical modifiability, extremely high photostability and biocompatibility, and provides a variety of physical information quantitatively via optically detected magnetic resonance (ODMR) measurements. We performed the chemical surface modification of an ND with the actin filament-specific binding peptide Lifeact and observed colocalization of pure Lifeact-modified ND and actin filaments by the ODMR selective imaging protocol, suggesting the capability of long-term observation and quantitative analysis of a single molecule by using an ND particle.