使用自适应相位补偿超短激光脉冲的双光子成像。

Two-photon imaging using adaptive phase compensated ultrashort laser pulses.

作者信息

Xi Peng, Andegeko Yair, Pestov Dmitry, Lovozoy Vadim V, Dantus Marcos

机构信息

Shanghai Jiao Tong University, Department of Biomedical Engineering, Institute for Laser Medicine and Biophotonics, Shanghai 200240 China.

出版信息

J Biomed Opt. 2009 Jan-Feb;14(1):014002. doi: 10.1117/1.3059629.

DOI:10.1117/1.3059629

PMID:19256690

Abstract

An adaptive pulse shaper controlled by multiphoton intrapulse interference phase scanning (MIIPS) was used, together with a prism-pair, to measure and cancel high-order phase distortions introduced by a high-numerical-aperture objective and other dispersive elements of a two-photon laser-scanning microscope. The delivery of broad-bandwidth (approximately 100 nm), sub-12-fs pulses was confirmed by interferometric autocorrelation measurements at the focal plane. A comparison of two-photon imaging with transform-limited and second-order-dispersion compensated laser pulses of the same energy showed a 6-to-11-fold improvement in the two-photon excitation fluorescence signal when applied to cells and tissue, and up to a 19-fold improvement in the second harmonic generation signal from a rat tendon specimen.

摘要

采用由多光子脉冲内干涉相位扫描（MIIPS）控制的自适应脉冲整形器，与一对棱镜一起，来测量和消除由高数值孔径物镜及双光子激光扫描显微镜的其他色散元件引入的高阶相位畸变。通过焦平面处的干涉自相关测量证实了宽带宽（约100 nm）、亚12飞秒脉冲的传输。对具有相同能量的变换极限和二阶色散补偿激光脉冲进行双光子成像比较，结果表明，当应用于细胞和组织时，双光子激发荧光信号提高了6至11倍，而来自大鼠肌腱标本的二次谐波信号提高了19倍。