Periodic acceptor excitation spectroscopy of single molecules
Periodic acceptor excitation spectroscopy of single molecules
复制标题
DOI：
10.1007/s00249-007-0133-7
复制
发表时间：
2007-07-01
影响因子：
2
通讯作者：
Kapanidis, Achillefs N.
中科院分区：
文献类型：
--
作者：
Doose, Soren;Heilemann, Mike;Kapanidis, Achillefs N.
交流激光激发（ALEX）光谱学最近被添加到单分子光谱学工具箱中。ALEX监测生物分子的相互作用和化学计量学，通过测量精确的福斯特共振能量转移（FRET）效率来报告生物分子结构，并允许在化学计量学和FRET的基础上对亚群进行分类。在这里，我们证明了一个连续波给体激发激光器和一个直接调制的受体激发激光器（周期性受体激发）的简单组合足以概括ALEX的能力，同时最大限度地降低与使用调制技术相关的成本和复杂性。
Alternating-laser excitation (ALEX) spectroscopy has recently been added to the single-molecule spectroscopy toolkit. ALEX monitors interaction and stoichiometry of biomolecules, reports on biomolecular structure by measuring accurate Forster resonance energy transfer (FRET) efficiencies, and allows sorting of subpopulations on the basis of stoichiometry and FRET. Here, we demonstrate that a simple combination of one continuous-wave donor-excitation laser and one directly modulated acceptor-excitation laser (Periodic Acceptor eXcitation) is sufficient to recapitulate the capabilities of ALEX while minimizing the cost and complexity associated with use of modulation techniques.