Anti-claudin-4, whose corresponding antigen receptors are known to be overexpressed in both primary and metastatic human pancreatic cancer, is utilized for targeted delivery and imaging of pancreatic cancer. In this protocol, we describe the use of quantum dots (QDs) as sensitive optical contrast agent for imaging pancreatic cancer in vitro and in vivo by using anti-claudin-4 as targeting ligands. The claudin-4-mediated targeting is demonstrated in using both in vitro confocal microscopy and in vivo tumor imaging system. This targeted QD platform will be further modified for the purpose of developing as an early detection imaging tool for pancreatic cancer.