使用线激发的16通道时间分辨单分子光谱学
16-Ch Time-resolved Single-Molecule Spectroscopy Using Line Excitation.
作者信息
Ingargiola Antonino, Peronio Pietro, Lerner Eitan, Gulinatti Angelo, Rech Ivan, Ghioni Massimo, Weiss Shimon, Michalet Xavier
机构信息
Dept. Chemistry & Biochemistry, University of California Los Angeles, Los Angeles, CA, USA.
DEIB, Politecnico di Milano, Milan, Italy.
出版信息
Proc SPIE Int Soc Opt Eng. 2017 Jan 28;10071. doi: 10.1117/12.2256367. Epub 2017 Mar 2.
Single-molecule spectroscopy on freely-diffusing molecules allows detecting conformational changes of biomolecules without perturbation from surface immobilization. Resolving fluorescence lifetimes increases the sensitivity in detecting conformational changes and overcomes artifacts common in intensity-based measurements. Common to all freely-diffusing techniques, however, are the long acquisition times. We report a time-resolved multispot system employing a 16-channel SPAD array and TCSPC electronics, which overcomes the throughput issue. Excitation is obtained by shaping a 532 nm pulsed laser into a line, matching the linear SPAD array geometry. We show that the line-excitation is a robust and cost-effective approach to implement multispot systems based on linear detector arrays.
对自由扩散分子进行单分子光谱分析，能够在不受表面固定化干扰的情况下检测生物分子的构象变化。解析荧光寿命可提高检测构象变化的灵敏度，并克服基于强度测量中常见的伪像。然而，所有自由扩散技术的共同问题是采集时间长。我们报道了一种采用16通道单光子雪崩二极管（SPAD）阵列和时间相关单光子计数（TCSPC）电子设备的时间分辨多光斑系统，该系统克服了通量问题。通过将532 nm脉冲激光整形为一条线来实现激发，使其与线性SPAD阵列的几何形状相匹配。我们表明，线激发是一种基于线性探测器阵列实现多光斑系统的稳健且经济高效的方法。
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