AbstractBackground and objectivesPhotoacoustic lymphangiography, which is based on photoacoustic technology, is an optical imaging that visualizes the distribution of light absorbing tissue components like hemoglobin or melanin, as well as optical absorption contrast imaging agents like indocyanine green (ICG) in the lymphatic channels, with high spatial resolution. In this report, we introduce the three‐dimensional (3D) images of human lymphatic vessels obtained with photoacoustic lymphangiography.MethodsWe used the 3D photoacoustic visualization system (PAI‐05). Some healthy subjects and lymphedema patients were recruited. To image the lymphatic structures of the limbs ICG was administered subcutaneously as in fluorescence lymphangiography. Photoacoustic images were acquired by irradiating the tissue using a laser at wavelengths of near‐infrared region. On the same occasion, fluorescence images were also recorded.ResultsThe lymphatic vessels up to the diameter of 0.2 mm could be observed three‐dimensionally with the venules around them. In the patient‐group, dermal backflow patterns were often observed as dense interconnecting 3D structures of lymphatic vessels. Collecting vessels passing below the dermis were also observed, which were not observed by fluorescence lymphography.ConclusionsPhotoacoustic lymphangiography provided the detailed observation of each lymphatic vessel, leading to deeper understanding of 3D structures and physiological state of the vessel.