Surface preparation is very important for the adhesion of thermal sprayed coatings. In this work, the effect of surface preparation on the adhesion strength of suspension plasma sprayed (SPS) Al2O3 coatings is investigated. Aluminum (Al-2024) substrates were micromachined by femtosecond laser (FSL) and also mechanically blasted by conventional grit blasting for surface preparation prior to SPS coating. Surface morphology results showed laser-induced periodic surface structures (LIPSS) produced by FSL, whereas grit blasting produced random irregularities with peaks and undercuts. Surface chemistry analysis showed that the FSL-prepared substrates were functionalized by -OH and O, while the grit-blasted substrates were not functionalized. The prepared substrates were coated by the SPS process under two spray conditions, where one condition was selected to produce a dense coating and the other condition was selected to produce a porous coating. The dense coating showed poor adhesion regardless of surface preparation, while the porous coating showed good adhesion for both grit-blasted and FSL-treated substrates. The adhesion strength of the porous coating was evaluated using the ASTM C-633 pull test protocol. A significant improvement (4.5 factors higher) in adhesion strength was achieved for the FSL-prepared substrates compared to the grit-blasted substrates. The improved adhesion of the FSL-prepared substrates was assessed by surface chemistry and surface morphology analyses of the prepared substrates, as well as by further analysis of the failed samples after the adhesion test and splat analysis. Results from this research showed that FSL has potential as an effective surface preparation technique for SPS.