A Nanodiamond-peptide Bioconjugate for Fluorescence and ODMR Microscopy of a Single Actin Filament.
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.
Department of Molecular Engineering, Graduate School of Engineering, Kyoto University.
Takuya
T
Genjo
Shingo
S
Sotoma
Ryotaro
R
Tanabe
Ryuji
R
Igarashi
Masahiro
M
Shirakawa
D008841:Actin Cytoskeleton / Q000737:chemistry*; D000199:Actins / Q000737:chemistry*; D000818:Animals; D001672:Biocompatible Materials; D001696:Biomechanical Phenomena; D003599:Cytoskeleton / Q000378:metabolism; D005453:Fluorescence; D008856:Microscopy, Fluorescence / Q000379:methods*; D018482:Muscle, Skeletal / Q000378:metabolism; D058612:Nanodiamonds; D009584:Nitrogen; D055095:Optics and Photonics; D010455:Peptides / Q000737:chemistry; D010590:Phalloidine / Q000737:chemistry; D011485:Protein Binding; D011817:Rabbits; D012235:Rhodamines / Q000737:chemistry
D000199:Actins; D001672:Biocompatible Materials; D058612:Nanodiamonds; D010455:Peptides; D012235:Rhodamines; D010590:Phalloidine; D009584:Nitrogen