Lane Richard S K, Macpherson Alisdair N, Magennis Steven W
The School of Chemistry, The University of Manchester, Oxford Road, Manchester, M13 9PL, UK.
Opt Express. 2012 Nov 5;20(23):25948-59. doi: 10.1364/OE.20.025948.
We demonstrate that pulse shaping of a broadband Ti:sapphire laser can result in almost an order of magnitude increase in the sensitivity and signal to background ratio (SBR) of multiphoton total internal reflection fluorescence (TIRF) microscopy. We produced transform-limited pulses of 15 fs duration at the sample, and observed a 8-fold enhancement in the fluorescence of CdSe/ZnS quantum dots via two-photon objective-type TIRF excitation. There was a concomitant 6-fold increase of the SBR upon compression of the pulse duration. Enhancement of non-linear evanescent imaging has recently been demonstrated using surface-plasmons [Opt. Express 17, 5987 (2009)] and structured substrates [Opt. Express 18, 23218 (2010)]. Our approach of ultrafast pulse shaping could be used alone or combined with these new methods to offer significant gains in image quality.
我们证明，宽带钛宝石激光器的脉冲整形可使多光子全内反射荧光（TIRF）显微镜的灵敏度和信号背景比（SBR）提高近一个数量级。我们在样品处产生了持续时间为15 fs的变换极限脉冲，并通过双光子物镜型TIRF激发观察到CdSe/ZnS量子点的荧光增强了8倍。在压缩脉冲持续时间时，SBR随之增加了6倍。最近已证明使用表面等离子体激元[《光学快报》17, 5987 (2009)]和结构化衬底[《光学快报》18, 23218 (2010)]可增强非线性倏逝成像。我们的超快脉冲整形方法可单独使用，或与这些新方法结合使用，以显著提高图像质量。
