Optimization of Intraoperative Near-Infrared Fluorescence Mapping with Indocyanine Green for Sentinel Lymph Node Detection in Cervical and Endometrial Cancer

<b>Background/Objectives:</b> Lymph node dissection during surgeries for cervical and endometrial cancer is associated with significant complications and morbidity. Sentinel lymph nodes (SLNs) mapping using indocyanine green (ICG) has become a promising method for reducing surgical invasiveness and improving patient outcomes. However, the optimal protocol for intraoperative fluorescence mapping of SLNs using ICG, especially regarding the timing of imaging after injection, remains to be fully optimized. This study aimed to evaluate the efficacy of real-time near-infrared (NIR) fluorescence SLN mapping at various time intervals and to investigate the photophysical properties of ICG in human lymph to establish a correlation between fluorescence signals and dye concentration. <b>Methods:</b> A prospective study included 20 patients with cervical and endometrial cancer undergoing laparoscopic or laparotomic surgery. Interstitial ICG injection was administered into the cervical stroma. SLN mapping was conducted using the novel VENERA-green endoscopic system (λ_exc = 800 nm, registration of fluorescence in the range of 830-1000 nm). Spectral fluorescence analysis (λ_exc = 650 nm) was conducted on SLNs and optical phantoms containing human lymph with ICG concentrations from 0 to 40 mg/L. The method made it possible to evaluate ICG absorption/emission properties, as well as to quantify concentration-dependent effects. <b>Results:</b> SLNs were successfully detected in all patients. The average detection time was 15 min with a range of 10 to 25 min. Fluorescence intensity of SLNs was significantly higher 10-15 min after ICG injection compared to 20-25 min. Spectral analysis indicated an absorption peak at 804 nm and an emission peak in the 835-855 nm range for ICG in human lymph. A concentration-dependent redshift of the fluorescence peak was observed and accurately modeled using a logarithmic function (R² = 0.99), which allows for the estimation of ICG concentration in tissue. The bilateral detection rate was 77% for laparoscopy and 100% for laparotomy. Metastases were histologically confirmed in only 2.8% (1/36) of the detected SLNs. <b>Conclusions:</b> Intraoperative NIR fluorescence imaging using ICG is a highly sensitive method for real-time SLN mapping in gynecologic oncology. The optimal detection period is 10 to 15 min after cervical injection to achieve maximum ICG fluorescence intensity, compared to 20 to 25 min. The concentration-dependent fluorescence and absorption properties of ICG in lymph provide the basis for the development of quantitative intraoperative monitoring methods that could improve the accuracy of sentinel lymph node biopsy.