The present invention presents a device and methods of use thereof in combined electrohydrodynamic concentration and plasmonic detection of a charged species of interest using a flow-through nanohole array. The device comprises microchannels, which are linked to a substrate with arrays of through nanoholes, wherein the substrate comprises two layers, wherein one of the layers is made of insulator material and one of the layers is made of metal, whereby induction of an electric field across the nanohole array results in the species of interest concentrating inside the nanoholes and in the vicinity of the nanohole arrays. The induction of an electric field is achieved by means of an external electric field source, which is applied to the fluid containing the species of interest, resulting in electroosmotic (EO) flow. An additional pressure driven fluid flow in the microchannels, co-directional to the EO flow is applied by external means. The resulting fluid flow from the combination of the EO and pressure driven flow results in a total bulk fluid flow hereafter referred to as bulk flow (BF). The local electric field strength across the insulator layer of the nanoholes is high and the charged species in the fluid may exhibit a high electrophoretic (EP) velocity, opposing the BF. The local field strength in the metallic portion of the nanoholes is null, due to the conducting nature of the metal, and the charged species in the fluid exhibits a null EP velocity in this region. The BF and the EP velocity of the charged species may be balanced which may result in the concentration of the charged species inside the nanoholes and at both sides of the nanohole array. An incident light over one side of the nanohole array may result in the formation of surface plasmons (SP) at the interface of the metal and the surrounding liquid containing the concentrated species. The signal from the SP may be detected by optical means, including surface plasmon resonance (SPR) imaging and SPR spectroscopy.