Carbocyanine dyes as efficient reversible single-molecule optical switch
Carbocyanine dyes as efficient reversible single-molecule optical switch
复制标题
DOI：
10.1021/ja044686x
复制
发表时间：
2005-03-23
影响因子：
15
通讯作者：
Tinnefeld, P
中科院分区：
文献类型：
--
作者：
Heilemann, M;Margeat, E;Tinnefeld, P
我们证明了市售的未经修饰的碳菁染料如Cy5（通常在633 nm激发）可以用作有效的可逆单分子光开关，其在明显光漂白后的荧光状态可以在室温下在较短波长的照射下恢复。Cy5在633 nm、337、488或532 nm辐照的水溶液中进行的整体光漂白和回收实验表明，吸收和荧光的恢复强烈依赖于有效的除氧和三重态猝灭剂β -巯基乙胺的加入。单分子荧光实验表明，在室温下，在633和488 nm处交替激发荧光和非荧光状态100次，单个固定的Cy5分子可以在几毫秒内进行光学切换，可靠性为> 90%。由于其令人感兴趣的性能，碳菁染料自愿成为超高密度光学数据存储的简单替代方案。对单个供体/受体（四甲基罗丹明/Cy5）标记的寡核苷酸的测量指出，所描述的光驱动开关行为对使用碳菁染料作为生物过程研究的能量转移受体施加了根本性的限制。
We demonstrate that commercially available unmodified carbocyanine dyes such as Cy5 (usually excited at 633 nm) can be used as efficient reversible single-molecule optical switch, whose fluorescent state after apparent photobleaching can be restored at room temperature upon irradiation at shorter wavelengths. Ensemble photobleaching and recovery experiments of Cy5 in aqueous solution irradiating first at 633 nm, then at 337, 488, or 532 nm, demonstrate that restoration of absorption and fluorescence strongly depends on efficient oxygen removal and the addition of the triplet quencher beta-mercaptoethylamine. Single-molecule fluorescence experiments show that individual immobilized Cy5 molecules can be switched optically in milliseconds by applying alternating excitation at 633 and 488 nm between a fluorescent and nonfluorescent state up to 100 times with a reliability of > 90% at room temperature. Because of their intriguing performance, carbocyanine dyes volunteer as a simple alternative for ultrahigh-density optical data storage. Measurements on single donor/acceptor (tetramethylrhodamine/Cy5) labeled oligonucleotides point out that the described light-driven switching behavior imposes fundamental limitations on the use of carbocyanine dyes as energy transfer acceptors for the study of biological processes.