In this work, the theory presented in part 1 (van der Veen, M. A.; Vermoortele, F.; De Vos, D. E.; Verbiest, T. Anal. Chem. 2012, DOI: 10.1021/ac300936q) for determination of the point groups symmetry based on easily distinguishable observables present in simple polarization dependent tests in second harmonic generation microscopy is tested. It is shown experimentally that the methodology can be applied for point group symmetry determination for a variety of structures among which molecular crystals and host/guest systems where the symmetry of the guest molecules cannot be inferred from conventional diffraction methods. Uniquely, this second-harmonic generation based method can discriminate between chiral and achiral structures regardless of their orientation. The method allows for in situ and in vivo studies with spatial resolution.