Aim: The objective of this study was to compare indocyanine green (ICG) and blue colorimetric methods for detection of sentinel lymph nodes (SLN) during laparoscopic surgery for uterine malignancies. Materials and Methods: Over a two years and 3months period, laparoscopic surgery was performed on 16 cases of uterine malignancies using near infrared (NIR) fluorescence imaging of ICG tracer uptake to direct SLN biopsy (NIR-FI-SLNB). ICG was injected into the uterine cervix prior to surgery. For comparison of detection efficacy, blue dye used for traditional colorimetric method was injected concurrently into 14 of these 16 cases. After pneumoperitoneum, we opened the retroperitoneum to laparoscopically identify the SLN. After biopsy of the SLN, a systemic pelvic lymph node dissection was performed. Results: Using ICG, we were able to identify SLN in 15/16 cases (93.7%), and achieved bilateral pelvic mapping in 12/16 (75%). With the blue dye, we were also able to identify SLN in only 64.3% of the 14 cases so tested, and bilateral pelvic mapping in only 14.3%. There were no intraoperative complications during any of the procedures. For conducting a laparoscopic bilateral SLN biopsy, we found that use of ICG was significantly superior to the blue colorimetric method (14.3% vs. 75%, p = 0.0009). Conclusions: These pilot data provide suggestive evidence that the laparoscopic NIR fluorescence imaging for SLN biopsy provide superior efficacy, compared to the traditional blue colorimetric method, without additional complications.