The preparation of ultra flat gold surfaces for use in chemical force microscopy (CFM) has been studied. The surfaces were studied in terms of substrate effects by comparing mica, Si (110) wafer and glass slides. The effect of different annealing regimes was also investigated. Measurements on these surfaces were made by both atomic force microscopy (AFM) (in contact and tapping mode) and by scanning tunneling microscopy (STM). The films contain different morphologies with respect to grain size and topography. Calculations of surface roughness present values less than 2.5 nm for all surfaces studied, making the choice of the flattest surface difficult if based on criteria of surface roughness alone. Additionally, it is shown that different acquisition parameters can produce dissimilar images that have stability and reproducibility.