We developed second-generation (G2) optical-resolution photoacoustic microscopy (OR-PAM). Incorporation of a novel acoustic detection scheme improved upon the sensitivity of our first-generation (G1) system by 18.4 dB, deepening the in vivo tissue penetration to 1.2 mm at 570 nm. Moreover, translating the imaging head instead of the living object accelerated the scanning speed by a factor of 5, widening the field of view within the same acquisition time. Mouse ears, as well as mouse brains with intact craniums, were imaged in vivo in both total concentration and oxygen saturation of hemoglobin.