Two-photon microscopy (TPM) is minimally-invasive 3D fluorescence microscopy based on nonlinear excitation, and TPM can visualize cellular structures based on auto-fluorescence. Line-scanning TPM is one of high-speed TPM methods without sacrificing the image resolution by using spatial and temporal focusing. In this paper, we developed line-scanning TPM based on spatial and temporal focusing for auto-fluorescence imaging by exciting the tryptophan. Laser source for this system was an optical parametric oscillator (OPO) and it made near 570 nm femtosecond pulse laser. It had 200fs pulse width and 1.72 nm bandwidth, so that the achievable depth resolution was 2.41um and field of view (FOV) is 10.8um. From the characterization, our system has 3.0 um depth resolution and 12.3 um FOV. We visualized fixed leukocyte cell sample and compared with point scanning system.