Post-operative hypoparathyroidism still remains a significant complication after thyroidectomy. Intraoperative imaging modalities, such as near infrared fluorescence by the use of indocyanine green (ICG), may assist to identify and preserve the parathyroid glands (PGs). The purpose of this study was to test the association between intra-operative ICG staining scoring system and 24-h post-operative parathyroid hormone (PTH) levels, as well as its capability of intra-operative PG identification. This was a prospective study, recruiting patients scheduled for total thyroidectomy by the same surgical team, from December 2018 to April 2019. Intraoperative angiography was performed, after infusion of ICG solution (5 mg). Two minutes later, images were acquired by the near-infrared system. Sixty patients fulfilled eligibility criteria. The percentage of temporary post-operative hypoPT (defined as PTH <14 pg/mL) was 11.66%. No association between intra-operative ICG staining score (expressed as the number of PGs scoring <2 per patient) and 24-h post-operative PTH (r=0.011, =0.933) or serum calcium concentrations (r=0.127, =0.335). There was also no correlation between the location of PGs scoring ≤2 and post-operative PTH (=0.257) or serum calcium levels (=0.950). Moreover, with regard to secondary endpoint, ICG correctly identified parathyroid glands in 98.3% of cases. ICG score was not affected by age, gender, duration of operation or thyroid gland pathology. No allergic reactions attributed to ICG administration were observed. Intra-operative ICG staining scoring system did not predict 24-h post-operative PTH and serum calcium levels. However, this modality may assist in intra-operative parathyroid identification during a total thyroidectomy.