Understanding the structural dynamics underlying the biological actions of isatin-3 thiosemicarbazone and its derivatives has advanced significantly thanks to the conformational characterization of these compounds. In order to properly analyse the conformational preferences of these chemicals, this work uses a multimodal strategy that integrates both computational and experimental approaches. The potential energy surfaces are investigated, stable conformers are found, and comprehensive insights into the electronic structure are obtained by Density Functional Theory (DFT) computations. Nucleus magnetic resonance (NMR), infrared (IR), and ultraviolet-visible (UV-Vis) spectroscopy are among the experimental methods used to confirm the computational results and to provide a complete picture of the conformational landscape in various settings.