In this work, we investigated the principle of the two-photon absorption (TPA) detection with a loss modulation technique, and first demonstrated the existence of two-photon photoacoustics ultrasound excited by a femtosecond high repetition rate laser. By using the AO modulation with different modulation frequencies, we successfully create the beating of the light signal when the two arms of the beams are both spatial and temporal overlapping. The pulse train of the femtosecond laser causes the narrow band excitation, providing the frequency selectivity and sensitivity. Moreover, the pulse energy is no more than 15nJ/pulse, which is at least 3 orders of magnitude smaller than that of the nanosecond laser, and therefore prevents the thermal damage of the sample. With the help of lock-in detection and a low noise amplifier, we can separate the signal of two-photon absorption from one-photon absorption. We used an ultrasonic transducer to detect the response of the sample, and verified the existence of the two-photon photoacoustics ultrasound generating by the femtosecond laser. Several contrast agents, such as the black carbon solution, the fluorescence dye and the nano-particles, were used in the experiment. In the end, we demonstrated the application, two photo-acoustic imaging, which provides the high spatial resolution (<10μm) and large penetration depth (~1mm), to the simulated biological tissue. This is a milestone to develop the two-photon photoacoustics microscopy, which, in principle, has the great potential to achieve the in vitro and in vivo high resolution deep tissue imaging.