Although several intraoperative methods to localize parathyroid gland(PG) were introduced, their clinical application has been limited. Thus, the current solution has been based on visual inspection of the surgeon. Recently, we reported excellent results (PPV = 100%, n=16) of Near infrared and infrared imaging technique for identifying surgically exposed PGs (JCEM, 2016). However, it is more important to predict the location of PGs before they are exposed to the naked eye. We investigated the feasibility of PG mapping (navigation) with NIR autofluorescence imaging to localize unexposed PGs. Seventy PGs from 38 thyroidectomy patients were enrolled in this prospective study. NIR imaging was taken at the areas where PGs were predicted to exist. We named the procedure as Parathyroid Navigation. The parathyroid navigation was photographed in three stages. Stage P1, taking images at pre-identification by the naked eye, step P2, at post-identification, and step P3, at removed specimens. Of 69 PGs identified, 64 (92.8%) were found at P1, 4 (5.8%) at P2, and 1 (1.4%) at P3. Even when PGs were covered by connective or fat tissues, NIR navigation at step P1 showed the sensitivity (92.75 %), specificity (100 %), PPV (100 %), NPV (16.66 %), accuracy (92.85 %). Five PGs of step P1 negative were identified at step P2 and P3 revealing 100% of total accuracy rate. The average parathyroid/background ratio was 4.78. These results suggest the concept of parathyroid navigation is feasible. We believe surgeons can get benefits of preserving parathyroid gland with the use of our NIR imaging method.