By utilizing Dy(hfac)3(H2O)2to react with enantiomerically pure tridentate N,N,N-pincer ligands, namely (−)/(+)-2,6-bis(4′,5′-pinene-2′-pyridyl)pyridine (LRand LS), respectively, homochiral DyIIIenantiomeric pairs formulated as Dy(hfac)3LR/Dy(hfac)3LS(R-1/S-1) (hfac–= hexafluoroacetylacetonate) were achieved and structurally characterized. Meanwhile, their magnetic, photoluminescent (PL), and chiroptical properties were probed. The PL test results indicate that the precursor Dy(hfac)3(H2O)2only shows very weak emission, while R-1exhibits characteristic DyIIIf–f transition emission bands at room temperature. Furthermore, the nonlinear optical responses of Dy(hfac)3(H2O)2, LR/LS, and R-1/S-1were investigated in detail based on crystalline samples. The results reveal that LRand LSpresent the coexistence of second- and third-harmonic generation (SHG and THG) responses with more intense signals for SHG responses; and Dy(hfac)3(H2O)2merely displays weak THG responses, while R-1and S-1also only exhibit THG responses. However, the THG intensities of R-1and S-1are more than six times larger than that of Dy(hfac)3(H2O)2under the identical measurement conditions. These results demonstrate that introducing homochiral N,N,N-pincer ligands to replace two H2O molecules of Dy(hfac)3(H2O)2results in significant improvements of both PL performances and THG responses of resultant R-1/S-1enantiomers. R-1and S-1integrate PL, THG, and chiral optical activity in one molecule, suggesting their multifunctional merits. In particular, a convenient method is introduced to simultaneously test THG and SHG responses of molecular materials based on crystalline samples in this work.