Two-photon imaging using adaptive phase compensated ultrashort laser pulses

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

doi:10.1117/1.3059629

Two-photon imaging using adaptive phase compensated ultrashort laser pulses

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

Authors

Peng Xi¹, Yair Andegeko, Dmitry Pestov, Vadim V Lovozoy, Marcos Dantus

Affiliation

¹ Shanghai Jiao Tong University, Department of Biomedical Engineering, Institute for Laser Medicine and Biophotonics, Shanghai 200240 China.

PMID: 19256690
DOI: 10.1117/1.3059629

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.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Algorithms*
Equipment Design
Equipment Failure Analysis
Image Enhancement / instrumentation*
Image Enhancement / methods
Image Interpretation, Computer-Assisted / instrumentation*
Image Interpretation, Computer-Assisted / methods
Lasers*
Microscopy, Confocal / instrumentation*
Microscopy, Fluorescence, Multiphoton / instrumentation*
Microscopy, Fluorescence, Multiphoton / methods
Reproducibility of Results
Sensitivity and Specificity
Signal Processing, Computer-Assisted / instrumentation*