As a promising luminescent material, the two-dimensional (2D) perovskite exhibits efficient and tunable photoluminescence due to its naturally self-assembled multiple quantum well structure and consequently large exciton binding energy. [20] The structural stability of 2D perovskites over their three-dimensional (3D) counterpart, owing to the hydrophobic nature of organic barriers and the van der Waals interaction of neighboring layers, has motivated further studies to engineer these layered structures and enhance their properties. [7,13,21] The crystal growth of 2D perovskite is usually parallel to the substrate, leading to a preferential orientation of layered structures in the polycrystalline film. [22,23] The structural anisotropy brings in not only significant difference between in-plane and out-of-plane conductivities [24] but also the unique emission performance at different angle. [25] In this case, the grain boundaries could break the local structural symmetry for dipole transitions, [26][27][28] and influence the anisotropic property of photoluminescence through the emissive in-gap state. Moreover, the interplay between in-gap state and band-gap state provides possibilities in developing bi-directionally controlled light emission from 2D perovskite films.In this work, we investigated the in-gap state related to grain boundaries and observed an significant enhancement of photoluminescence anisotropy from the (PEA) 2 PbI 4 (PEA is phenethylamine) films by reducing the grain size via hotcasting method. [29] The preparation temperature can change the morphology of 2D perovskite films and thus control the density of in-gap state, which is characterized by measuring the photoconductivity as the carrier dissociation is facilitated by grain boundaries. The photoluminescence of in-gap state shows an ≈20 nm red-shift compared with that of band-gap state, and more interestingly, is spatially anisotropic meaning that the light intensity is strongly dependent on the collection angle. The grain boundaries are found to enhance the in-gap emission and consequently bring the photoluminescence anisotropy from (PEA) 2 PbI 4 films. The origin of photoluminescence anisotropy is that the transition dipole moment of in-gap state is perpendicular to that of band-gap state, revealed by measuring the light polarization of photoluminescence bands.The (PEA) 2 PbI 4 polycrystalline films were prepared by a spin-casting method (see details in Supporting Information). The substrate was pre-heated to facilitate the solvent evaporation and thus the crystallization process during spin-casting, asThe boundaries between crystalline grains in hybrid perovskite films are beneficial for carrier dissociation, and may also serve as recombination centers to alter the photoluminescence property. Herein, it is shown that the grain boundaries can significantly enhance the anisotropy of photoluminescence from two-dimensional (2D) hybrid perovskite films. The measurement of carrier densities shows that the grain boundaries generate an in-gap state, ...