Imaging of the fluorescence spectrum of a single fluorescent molecule by prism‐based spectroscopy

FEBS Letters

Suzuki, Yoshikazu²  Tani, Tomomi¹  Sutoh, Kazuo²  Kamimura, Shinji² 

[1]The Tokyo Metropolitan Institute of Medical Science, Honkomagome 3-18-22, Bunkyo, Tokyo 113-8613, Japan

[2]Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Komaba 3-8-1, Meguro, Tokyo 153-8902, Japan

关键词: Dispersion prism;    Single-molecule spectral imaging;    Single-pair fluorescence resonance energy transfer;    Fluorescence microscopy;    FRET;    fluorescence resonance energy transfer;    TIRFM;    total internal reflection fluorescence microscopy;    ELC;    essential light chain of myosin;    TEAB;    triethylamine bicarbonate buffer;    EDC;    1-ethyl-3-(3-dimethylaminopropyl)carbodiimide;   

DOI  :  10.1016/S0014-5793(02)02269-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have devised a novel method to visualize the fluorescence spectrum of a single fluorescent molecule using prism-based spectroscopy. Equiping a total internal reflection microscope with a newly designed wedge prism, we obtained a spectral image of a single rhodamine red molecule attached to an essential light chain of myosin. We also obtained a spectral image of single-pair fluorescence resonance energy transfer between rhodamine red and Cy5 in a double-labeled myosin motor domain. This method could become a useful tool to investigate the dynamic processes of biomolecules at the single-molecule level.

【 授权许可】

Unknown