Fluorescence correlation spectroscopy based upon two-photon excitation

Abstract

Fluorescence correlation spectroscopy (FCS) is a powerful tool for measurement of biological dynamic processes. In this studying, a two-photon excitation fluorescence correlation spectroscopy (TP-FCS) system was set up depending on a part of optical block and detector of the multi-photon excitation fluorescence microscope (MPLFM). The phenomenon "photon-burst" was observed from the TP-FCS system. Meanwhile, the diffusion coefficient of rhodamine B molecule in sucrose aqueous solution was calculated. It was proved that TP-FCS is especially suited for integration into MPEFM accordingly to yield a hybride-technology with the peculiarities of the individual technique and the advantage of mutual synergistic effects. The fusion of both techniques seems to be reasonable and desirable to reduce costs.