Sentinel lymph nodes (SLNs) are commonly identified using a combination of blue dye and radioisotope methods. The objective of this study was to evaluate the feasibility of performing SLN biopsies using a fluorescence method with indocyanine green (ICG) and a near-infrared camera in combination with conventional blue dye and radioisotope methods in patients with malignant melanoma of the skin. A total of 49 patients with cutaneous melanoma underwent an SLN biopsy using a combination of the three methods. In 2 of the patients, SLNs could not be identified using either lymphoscintigraphy or a hand-held gamma probe. Since fluorescence from the lymph nodes in these 2 patients was, however, observed from the skin surface using a near-infrared camera, the SLNs were successfully identified and resected. The detection rate of SLNs using a combination of blue dye, radioisotope and a fluorescence method was 100%. SLN biopsy guided by indocyanine green fluorescence is a promising procedure in patients with cutaneous melanoma, especially those with a poor flow of radioactive tracer substance. Because it is difficult to detect anatomically deep SLNs and residual SLNs after SLN biopsy using the fluorescence method alone, the combined use with other identification methods is advisable.