ABSTRACT: We have developed a new actuation method for matrix-assisted ionization with good temporal and spatial resolution using piezoelectric cantilever. A strike from the piezoelectric bimorph cantilever on a thin metal foil was used to remove materials deposited on the opposite side facing the mass spectrometer inlet. Highly charged ions of peptides and proteins were generated from dried droplet deposits and sampled into the inlet of the mass spectrometer. A lateral resolution of 1 mm was obtained with the piezoelectric sampling configuration. Singly charged lipids and gangliosides were detected from tissue with piezoelectric matrix-assisted ionization using a silica nanoparticle co-matrix.