Experimental studies in immunology, pharmacology, or hematology require the sampling of the total thoracic duct lymph in awake and unrestrained rats or mice. Several approaches have been described for cannulation of the thoracic duct, but they are characterized by a modest reproducibility and a low lymph flow rate. An improved technique for obtaining thoracic duct lymph is described here, emphasizing the similarities and differences concerning both rats and mice (average weights of 305 and 15 g, respectively). Rats yielded a mean of 55.6 ml/day thoracic duct lymph, while lymph output in mice reached unexpected volumes of 29.3 ml/day. The use of an operating microscope and silicone cannula, and maintenance of mobility of the animals during lymph collection, offer a reliable method for a high and constant output of thoracic duct lymph. Relevant aspects of the murine thoracic duct anatomy are also identified. Copyright 2003 Wiley-Liss, Inc.