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A quantitative understanding of the nanoscale piezoelectric property will unlock many application potentials of the electromechanical coupling phenomenon under quantum confinement. In this work, we present an atomic force microscopy- (AFM-) based approach to the quantification of the nanometer-scale piezoelectric property from single-crystalline zinc oxide nanosheets (NSs) with thicknesses ranging from 1 to 4 nm. By identifying the appropriate driving potential, we minimized the influences from electrostatic interactions and tip-sample coupling, and extrapolated the thickness-dependent piezoelectric coefficient ( d 33 ). By averaging the measured d 33 from NSs with the same number of unit cells in thickness, an intriguing tri-unit-cell relationship was observed. From NSs with 3 n unit cell thickness ( n = 1 , 2, 3), a bulk-like d 33 at a value of ~9 pm/V was obtained, whereas NSs with other thickness showed a ~30% higher d 33 of ~12 pm/V. Quantification of d 33 as a function of ZnO unit cell numbers offers a new experimental discovery toward nanoscale piezoelectricity from nonlayered materials that are piezoelectric in bulk.