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Two-dimensional Mie-scattering and laser-induced fluorescence techniques were applied to investigate the effects of fuel composition on mixture formation within a firing direct-injection spark-ignition (DISI) engine. A comparison was made between the spray characteristics and in-cylinder fuel distributions resulting from the use of a typical multi-component gasoline (European specification premium-grade unleaded), a single-component research fuel (iso-octane), and a three-component research fuel (iso-pentane, iso-octane and n-nonane). Studies were performed at three different injection timings under cold and part-warm conditions. The results indicate that fuel composition affects both the initial spray formation and the subsequent mixture formation process. Furthermore, the sensitivity of the mixing process to the effects of fuel volatility was shown to depend on injection timing.