Tinbergen was the brother of the economist Jan Tinbergen. After receiving a Ph.D. degree (1932) from the University of Leiden, he taught there until 1949. He then served on the faculty of the University of Oxford (1949–74), where he organized a research department of animal behaviour. He became a British citizen in 1955.

With Lorenz and Frisch, Tinbergen is credited with revitalizing the science of ethology. Their emphasis was on field observations of animals under natural conditions. Tinbergen emphasized the importance of both instinctive and learned behaviour to survival and used animal behaviour as a basis for speculations about the nature of human violence and aggression. He is especially well known for his long-term observations of sea gulls, which led to important generalizations on courtship and mating behaviour.

Among his more important writings are The Herring Gull's World (1953; rev. ed. 1961), Social Behavior in Animals (1953), and Animal Behavior (1965). Perhaps his most influential work is The Study of Instinct (1951), which explores the work of the European ethological school up to that time and attempts a synthesis with American ethology. In the 1970s Tinbergen devoted his time to the study of autism in children.