Multifocal Two-photon Excitation Fluorescence Sampling Imaging Combining Lifetime and Spectrum Resolutions

摘要

Multifocal multiphoton microscopy (MMM) is a more efficient and powerful method for three-dimensional (3-D) fluorescence imaging with reduced acquisition time compared with conventional confocal and two-photon excitation fluorescence microscopy. We present a novel multifocal two-photon excitation fluorescence sampling imaging technique that is based on a specially designed streak camera and combines fluorescence lifetime and spectrum resolutions. A proof-of-principle experiment is performed on a standard fluorescent dye solution (Rhodamine 6G in ethanol), Time- and spectrum-resolved sampled fluorescence image of Rhodamine 6G is obtained in a snapshot. The reconstructed two-dimensional (2-D) fluorescence image of a prepared plant slide is also obtained by moving the sample laterally. The capability of this system capable of performing simultaneous 2-D measurements of temporal and spectral information has many potential applications, e.g., multi-well imaging and spectrally resolved multifocal multiphoton fluorescence lifetime imaging etc.