Photoacoustic (PA) imaging is an emerging, non-invasive imaging modality that encompasses attributes of both optical and ultrasound imaging. Because of the combination of optical excitation and acoustic detection, PA imaging enables high contrast and high resolution within deep tissue (centimeter depths). Recent advances in PA probe development have allowed for stimulus-responsive imaging in a variety of biological models with implications for basic, translational, and clinical sciences. This perspective highlights recent progress in the development of PA probes and their application to live-animal molecular imaging.