With broadband femtosecond (FS) oscillator as the excitation light source, a two-photon fluorescence (TPF) microscopy imaging system was built. Under the broadband excitation of FS pulses, TPF spectrum of Rhodamine Bsolution was measured and TPF microscopy imaging study of solid Rhodamine Bsamples was performed. In the study of TPF microscopy, the relationship between TPF intensity and excitation pulses power was analyzed. Besides, the trend of the two-photon fluorescence intensity with the polarization direction of the excitation pulse was studied through changing the polarization direction of the linearly polarized excitation light field by using a half-wave plate, and the sinusoidal-like modulation of the two-photon fluorescence intensity was realized.