The cancellation of De Monts's fur monopoly in 1607 brought the Port Royal settlement to a temporary end. Champlain persuaded his leader to allow him to take colonists and "go and settle on the great River St. Lawrence, with which I was familiar through a voyage that I had made there." In 1608 he founded France's first permanent Canadian colony. It was at Quebec, at the foot of a great rocky cape on the north shore, which formed a natural fortress barring the way upstream to the interior.

The early years of the Quebec colony were hard, and the population grew slowly. Champlain administered its affairs and took personal charge of an organized exploration of the unknown interior. Where he did not actually travel himself, he sent other men. One was Etienne Brule, the first white man to cross Pennsylvania and later the first to see Lake Superior. Champlain himself discovered Lake Champlain (1609); and in 1615 he journeyed by canoe up the Ottawa, through Lake Nipissing, and down Georgian Bay to the heart of the Huron country, near Lake Simcoe. During these journeys Champlain aided the Hurons in battles against the Iroquois Confederacy. As a result, the Iroquois became mortal enemies of the French.

In 1629 Champlain suffered the humiliation of having to surrender his almost starving garrison to an English fleet that appeared before Quebec. He was taken to England as a prisoner. Peace, however, had been declared between England and France before the surrender, and New France was accordingly restored to the French. Champlain returned from Europe to spend his few remaining years. He became governor of New France in 1633.

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