Optical Coherence Tomography (OCT) is a new high spatial resolution, real-time optical imaging modality, known from prior pilot studies for its high sensitivity to invasive cancer. We reported our results in an OCT feasibility study for accurate determination of the proximal border for esophageal carcinoma and the distal border for rectal carcinoma. The OCT study enrolled 19 patients with rectal adenocarcinoma and 24 patients with distal esophageal carcinoma (14 squamous cell carcinomas, 10 adenocarcinomas). During pre-surgery planning endoscopy we performed in vivo OCT imaging of the tumor border at four dial clock axes (12, 3, 6 and 9 o’clock). The OCT border then was marked by an electrocoagulator, or by a methylene blue tattoo. A cold biopsy (from the esophagus) was performed at visual and OCT borders and compared with visual and OCT readings. 27 post-surgery excised specimens were analyzed. OCT borders matched the histopathology in 94% cases in the rectum and 83.3% in the esophagus. In the cases of a mismatch between the OCT and histology borders, a deep tumor invasion occurred in the muscle layer (esophagus, rectum). Because of its high sensitivity to mucosal cancer, OCT can be used for pre-surgery planning and surgery guidance of the proximal border for esophageal carcinoma and the distal border for rectal carcinoma. However, deep invasion in the rectum or esophageal wall has to be controlled by alternative diagnostic modalities.