This paper aimed to describe the use of a new technology, RUBINA™, to perform intra-operative ICG fluorescent cholangiography (FC) in pediatric LC. ICG-FC was performed during LC using the new technology RUBINA™ in two pediatric surgery units. The ICG dosage was 0.35 mg/Kg and the median timing of administration was 15.6 h prior to surgery. Patient baseline, intra-operative details, rate of biliary anatomy identification, utilization ease, and surgical outcomes were assessed. Results showed that the new RUBINA™ technology was very effective to perform ICG-FC during LC in pediatric patients allowing surgeons fluorescence imaging of the target anatomy to assess position of critical biliary structures or presence of anatomical anomalies to safely perform the operation.