Colon cancer (CC) kills by metastatic spread to locoregional lymph nodes (LN's) and distant organs. One of the major unresolved issues in CC biology is, whether lymphatic spread is a stepwise, orderly process (Halsted model) or a stochastic process, in which metastasis to nearby nodes, more distant nodes, and distant organs occur randomly (Fisher model). The surgical standard of care for CC is wide resection of the tumor and the associated LN bearing mesentery. An adequate LN count is important for accurate staging. Moreover, numerous studies have shown a correlation between the number of examined nodes and overall survival in CC. However, recent data suggest that extensive lymphadenectomy does not have any therapeutic value and therefore, a limited (segmental) resection may provide a less morbid alternative, provided adequate nodal staging is accomplished. The latter is possible by the use of sentinel lymph node (SLN) techniques, which consist of identifying the first draining tumor LN. This SLN is considered to represent the status (invaded or not) of the entire nodal basin. The recently introduced near infrared (NIR) fluorescent optical imaging method, which can be incorporated in laparoscopic imaging hardware, was shown to result in superior accuracy, image resolution, and clinical applicability compared to the more traditional dye or radioactivity based methods. The drainage of tumor-derived factors through the lymphatic system to the regional LN's plays an important role in the pre-metastatic conditioning of the nodal microenvironment, making them receptive and supportive metastatic niches for disseminating tumor cells. Modified immunological responses are amongst the most important tumor-induced pre-metastatic changes in the lymph node microenvironment. Fluorescence based NIR using a large tracer such as ICG-nanocoll allows not only to identify the SLN, but also to spatially map lymphatic spread in CC.