Combining biosensors with nanoscience provides great advantages such as being label-free and real time, highly sensitive, and small in size, as well as providing a low limit of detection and integration to other systems. That is why plasmonics finds various applications in drug detection, food safety, agriculture, photothermal therapy, etc. In this paper, we have fabricated a two-dimensional plasmonic grating biosensor using a soft lithography technique, which has eliminated some disadvantages of conventional plasmonic structures like expensive fabrication cost, inflexibility, and lack of mass production. On the other hand, we benefited from infrared neural stimulation for regulating membrane depolarization, which was based on photothermal mechanism and provided a contact-free and high spatial/temporal resolution. Eventually, the membrane depolarization of two different cell types of Hep G2 and mesenchymal stem cells cultured on two-dimensional plasmonic structure has been investigated under infrared neural stimulation. After preparing the soft plasmonic crystal, its reflection spectra and respective ellipsometry parameters were analyzed before and after cell culture with/without stimulation (near-infrared immune region ∼1450 nm). By comparing the obtained ellipsometry results for HEP G2 and mesenchymal stem cells, it is observed that the behavior of two cell types with respect to IR stimulation was the same as well as providing us the possibility of distinguishing the level of membrane depolarization under various stimulating frequencies. The strength of this integrated system for membrane depolarization detection has been shown experimentally, which can open new avenues toward neuroplasmonic application in the future.