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This article investigates combinatorial properties of non-ambiguous trees. These objects we define may be seen either as binary trees drawn on a grid with some constraints, or as a subset of the tree-like tableaux previously defined by Aval, Boussicault and Nadeau. The enumeration of non-ambiguous trees satisfying some additional constraints allows us to give elegant combinatorial proofs of identities due to Carlitz, and to Ehrenborg and Steingrímsson. We also provide a hook formula to count the number of non-ambiguous trees with a given underlying tree. Finally, we use non-ambiguous trees to describe a very natural bijection between parallelogram polyominoes and binary trees.