Abstract: | Nonlinear optical (NLO) responses of perovskite‐type nanostructures have a variety of potential applications owing to the highly efficient frequency conversion guaranteed by both the material itself and the nanometer‐scale configuration. KNbO3 (KN) nanoneedles have been identified as a promising NLO material because of the superior broadband frequency conversion efficiency, and if incident light is propagating in a direction perpendicular to the axis of a nanoneedle, then the phase‐matching constraint can be relaxed. Here, the second‐harmonic generation (SHG) and third‐harmonic generation (THG) responses of both individual and clustered KN nanoneedles are reported. Based on these results, a novel method is proposed for determining the optimal excitation wavelength for NLO imaging of several biological samples, with KN nanoneedles acting as NLO agents. The method is shown to provide the optical features in the focal plane and a more reliable estimation of the optimal excitation wavelength for deep‐tissue imaging. |