Using field observations and laboratory experiments we examined the role of temperature in microhabitat selection by an eastern Ontario population of northern water snakes (Nerodia sipedon). From 1349 random transects through a marsh we found that basking activity peaked at 09:00 and then declined steadily until 14:00 before increasing again. Our ability to detect snakes depended upon the microhabitat they occupied, and to the time of day when the snakes were encountered in water. In the field, temperatures of basking snakes averaged (±SE) 26.3 ± 0.7 °C (n = 36), while captive snakes in a thermal gradient showed a narrower selectivity, averaging 27.7 ± 0.4 °C (n = 21). The temperatures of basking snakes never exceeded 33 °C, even though a model snake placed in the sun reached 48 °C, suggesting that the snakes were thermoregulating to prevent overheating. In both the field and enclosures, water snakes basked more frequently as the temperature of the air increased relative to the water. Experimental manipulation of water temperature relative to air temperature revealed that temperature influenced microhabitat selection independently of circadian patterns. Finally, when in water, snakes tended to frequent habitats where leopard frogs (Rana pipiens), a common prey species, were most abundant, suggesting that prey distribution may also be an important component of water snake habitat selection.