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题名: Spectral identification of specific photophysics of Cy5 by means of ensemble and single molecule measurements
作者: Huang, ZX;  Ji, DM;  Wang, SF;  Xia, AD;  Koberling, F;  Patting, M;  Erdmann, R
刊名: JOURNAL OF PHYSICAL CHEMISTRY A
发表日期: 2006-01-12
DOI: 10.1021/jp0562936
卷: 110, 期:1, 页:45-50
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Physical ;  Physics, Atomic, Molecular & Chemical
研究领域[WOS]: Chemistry ;  Physics
英文摘要: The triplet-state characteristics of the Cy5 molecule related to trans-cis isomerization are investigated by means of ensemble and single molecule measurements. Cy5 has been used frequently in the past 10 years in single molecule spectroscopic applications, e.g., as a probe or fluorescence resonance energy transfer acceptor in large biomolecules. However, the unknown spectral properties of the triplet state and the lack of knowledge on the photoisomerization do not allow us to interpret precisely the unexpected single molecule behaviors. This limits the application of Cy5. The laser photolysis experiments demonstrate that the trans triplet state of Cy5 absorbs about 625 nm, the cis ground state absorbs about 690 nm, and the cis triplet state also absorbs about 690 nm. In other words, the T-1-T-n absorptions largely overlap the ground-state absorptions for both trans and cis isomers, respectively. Furthermore, the observation of the cis triplet state indicates an important isomerization pathway from the trans-S-1 state to the cis-T-1 state upon excitation. The detailed spectra presented in this article let us clearly interpret the exact mechanisms responsible for several important and unexpected photophysical behaviors of single Cy5 molecules such as reverse intersystem crossing (RISC), the observation of dim states with a lower emission intensity and slightly red-shifted fluorescence, and unusual energy transfer from donor molecules to dark Cy5 molecules acting as acceptors in single molecule fluorescence resonance energy transfer (FRET) measurements. Spectral results show that the dim state in the single molecule fluorescence intensity time traces originated from cis-Cy5 because of a lower excitation rate, resulting from the red-shifted ground-state absorption of cis-Cy5 compared to that of the trans-Cy5.
关键词[WOS]: RESONANCE ENERGY-TRANSFER ;  FLUORESCENCE CORRELATION SPECTROSCOPY ;  CYANINE DYE ;  ELECTRON-TRANSFER ;  ROOM-TEMPERATURE ;  THIACARBOCYANINE DYES ;  BACK-ISOMERIZATION ;  TRIPLET-STATE ;  DYNAMICS ;  PHOTOISOMERIZATION
语种: 英语
WOS记录号: WOS:000234520300008
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/140008
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.PicoQuant GmbH, D-12489 Berlin, Germany
2.Chinese Acad Sci, Inst Chem, State Key Lab Mol React Dynam, Beijing 100080, Peoples R China

Recommended Citation:
Huang, ZX,Ji, DM,Wang, SF,et al. Spectral identification of specific photophysics of Cy5 by means of ensemble and single molecule measurements[J]. JOURNAL OF PHYSICAL CHEMISTRY A,2006,110(1):45-50.
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