It is demonstrated that a good biomimetic model lipid membrane with dynamic fluidity can be established on optically transparent nanocrystalline diamond (OTND) with surface roughness below 10 nm. Maigainin II, an antimicrobial peptide, is chosen to investigate the permeation of artificial bacterial membranes constructed on OTND. Due to the unique combination of optical transparency and highly sensitive surface conducting channel, intrinsic OTND affords the possibility of dual-mode sensing based on optical and field effect properties. This opens up new possibilities for making integrated biomolecule–semiconductor microdevices, or sensors where the binding of biomolecules can be tracked using confocal microscopy whilst the associated changes in charge density during membrane perforation can be tracked using the space charge effect in the semiconductor. Such a synergistic approach may provide a powerful methodology for the screening of specific bactericidal activity on biomimetic membrane systems.