These ISFETs were fabricated at the Institute of Electron Technology (IET) in Poland. A thermally grown SiO2 layer was deposited after RCA cleaning. Afterwards, the Si3N4 layer, a sensing membrane, was deposited by low pressure chemical vapor deposition (LPCVD). The gate width and length of the transistor channel are 600 ��m and 16 ��m, respectively. To aid in handmade encapsulation, extended source and drain areas with contact pads located away from the gate area were designed. Finally, all ISFETs were assembled on printed circuit boards (PCB) with a silver paste (TED PELLA, Inc.) and then encapsulated with epoxy resin type adhesive JU-100 (KOKI Company Ltd.) with open windows of 3 �� 3 mm2.2.3. Optimization of the PVC cocktail for REFETsIn order to decrease the pH sensitivity of the ISFET for REFET application, the PVC membranes were deposited on the open gate windows of ISFETs.

The fabrication process flow for the PVC membrane is illustrated in Figure 1. First, the gate insulator surface must be cleaned with deionized water and methanol. Then, for the purpose of chemical grafting and enhancing the adhesion between the PVC membrane and the Si3N4 layer, a silylating process based on hexamethyldisilazane (HMDS) deposited under different conditions is applied. Then, the PVC cocktail is cast on the Si3N4 surface of the ISFET with a micro pipette. The solvent from the PVC membrane is evaporated at room temperature overnight.Figure 1.The fabrication process flow for a REFET based on a Si3N4-ISFET.The procedure for REFET preparation was optimized by tuning the silylating process and PVC membrane composition.

The stability and adhesion of the PVC membrane depends mainly on the silylating process; therefore, four silylating processes for the Si3N4 layer with various HMDS treatments were investigated. The first samples were fabricated with a stock HMDS deposited directly and then dried at room temperature for 15 min, while the second batch of samples was baked at 120��C for 5 min. For the third method, a standard HMDS evaporation process that is used in photolithography was performed at 140��C in a vapor prime oven for 2 min. In the fourth method, HMDS diluted in toluene (ratio = 1:3) was deposited on the surface Drug_discovery of the Si3N4 layer and then dried at room temperature for 15 min.To prepare the PVC cocktails, three kinds of plasticizers with fixed weight percentage of 70 % were used: o-NPOE, DOS, and DNP. In addition, the content of the DNP plasticizer in the PVC cocktail was varied from 50 % to 80 % by weight vs. (PVC + DNP). For each batch of the PVC cocktail, six REFET samples were prepared. The total weight of the PVC + DNP was maintained at 200 mg, and all compounds were then dissolved in 3 mL of THF.2.4.