Objective This study opens a new direction for photoacoustic tomography (PAT) to image human cervical trachea.Methods A multi-wavelength PAT experiment was performed on the cervical trachea of seven volunteers. The characteristics of high resolution (up to 150 μm) images of the respiratory cycle and different layers of tracheal wall were studied by PAT, and the imaging advantages and defects of this technique were evaluated.Results Our study demonstrated the ability of PAT to image human cervical trachea, and delineate the structural characteristics of it and its surrounding tissues. Variation of tracheal lumen at different phases of a respiratory cycle by PAT indicated its high time resolution. Collagen, as an endogenous contrast of PAT, was abundant in tracheal cartilage, which has a wavelength of absorption coefficient ranging from 900 nm to 1 300 nm. This study shows that the wavelength of 910 nm is best for tracheal cartilage imaging.Conclusion It can be considered as a non-invasive, convenient and non-ionizing radiation imaging method for real-time evaluation of human trachea. Given the high spatial resolution and real-time capabilities, PAT has the potential to become a useful tool for non-invasive airway imaging in future. We believe this study sets the ground work for the analysis of the human trachea using PAT in vivo.