Estuarine migration of anosmic and control silver-phase American eels was examined during the fall spawning migration. Ultrasonic telemetry was used to track seventeen control and eight anosmic silver eels through 32 km of the Penobscot Estuary, Maine, U.S.A. Twelve of seventeen control eels migrated out of the estuary in 97 h (approximately 4 d) on average. Only two of eight anosmic eels migrated out of the estuary. On average these two anosmic eels migrated out of the estuary within 180 h (approximately 7.5 d) of release and the other six had not left within 9 d. Most control eels progressed rapidly to the mouth of the estuary within a few days. Anosmic eels spent more time in the estuary and demonstrated different behavior from control eels due to their lack of olfaction. Some control eels moved with the appropriate tide, the ebb tide for transport out of the estuary, within one tidal cycle of being released into tidal freshwater. However, anosmic eels either did not move with the appropriate tide or took significantly longer to do so. Olfaction was probably used for orientation by control eels sensing chemical clues (organic and inorganic), which change throughout the tidal cycle. Increased migration times and errors in orientation were likely related to the inability of anosmic eels to use selective tidal stream transport for movement out of the estuary. Chemical clues seem to be one of the most important environmental clues used to guide estuarine migration of silver eels. However, a hierarchy of sensory mechanisms and environmental clues are most likely used for estuarine orientation.