The spread of perfluoropolyether (PFPE) droplets on solid surfaces has been measured from the top-down view through a microscope system. Effects of substrates, molecular weight and end-group functionality on spreading of the PFPE droplets have been studied experimentally and the results were compared with those by molecular dynamics (MD)simulations. Silicon wafer and diamond-like carbon (DLC) substrates were used to study the effect of substrates on spreading. Two types of PFPE, Z-dol and Z-tetraol, with the same chain structure and various molecular weights (2000 and 4000 g/mol) were employed in experiments. Effect of molecular weight has been investigated through comparing the spreading of Z-dol 2000 and Z-dol 4000, and it is found that the increase of molecular weight will decrease the mobility of PFPE. Comparison between spreading of Z-dol and Z-tetraol of the same molecular weight proved that functional end group plays a significant role on the spreading of PFPE, which confirmed the MD simulation results.