One of the most crucial issues in the domain of microfluidics is the chip to world interface. This paper describes a characterization methodology of a quite common microfluidic interconnection scheme, based on polydimethylsiloxane (PDMS), applied to some of the most popular substrates (silicon, Pyrex and cyclic olefin copolymer) for microfluidic applications. Particular emphasis is given to the evaluation of leakage endurance as a function of the main geometrical parameters of the interconnections and the selected bonding technique. Oxygen plasma activation of the PDMS surface and the application of a thin PDMS interlayer demonstrated the most attractive solutions, due to the straightforward approach and limited cost. Maximum sustainable pressures in excess of 200 kPa have been achieved. Results obtained are critically discussed with the aim to outline PDMS interconnection guidelines for different microfluidic applications.