Abstract

Fluorescent proteins have been imperative tools to visualize nerve action or dynamic image of proteins in a living cell. Additionally two-photon excited fluorescence (TPEF) microscopy is a powerful tool for imaging proteins in a living cell because of its advantages such as high spatial resolution, three-dimensional imaging and reduced out-of-focus photobreaching. Especially, multicolor imaging and fluorescence resonance energy transfer (FRET) imaging are important techniques for investigating detailed cellular and molecular dynamics or functionality. We have developed a system for the nonlinear spectroscopic measurement of fluorescence proteins. In this paper, we present the two-photon excitation (TPE) spectra of various fluorescent proteins measured using Fourier-transform nonlinear spectroscopy with a spectral resolution of 60 cm⁻¹. By the use of an ultraboradband pulse with a spectrum ranging from 700 to 1100 nm, the TPE spectra ranging from 350 to 550 nm are measured.

© 2009 IEEE