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 B solution was measured and TPF microscopy imaging study of solid Rhodamine B samples 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.