Metal–semiconductor field effect transistors (MESFETs) based on hydrogen terminated diamond were fabricated according to different layouts. Aluminum gates were used on single crystal and low-roughness polycrystalline diamond substrates while gold was used for ohmic contacts. Hydrogen terminated layers were deeply investigated by means of Hall bars and transfer length structures. Room temperature Hall and field effect mobility values in excess of 100 cm2 V⁻¹ s⁻¹ were measured on commercial and single crystal epitaxial growth (100) plates by using the same hydrogenation process. Hydrogen induced two-dimensional hole gas resulted in sheet resistances essentially stable and repeatable depending on the substrate quality. Self-aligned 400 nm gate length FETs on single crystal substrates showed current density and transconductance values>100 mA mm⁻¹ and >40 mS mm⁻¹, respectively. Devices with gate length LG=200 nm showed fMax=26.4 GHz and fT=13.2 GHz whereas those fabricated on polycrystalline diamond, with the same gate geometry, exceeded fMax=23 GHz and fT=7 GHz. This work focused on the optimization of a self-aligned gate structure with respect to the fixed drain-to-source structure with which we observed higher frequency values; the new structure resulted in improvement of DC characteristics, better impedance matching and a reduction in the fMax/fT ratio.