In this study, we demonstrate a simple and efficient method to fabricate miniature surface plasmon resonance (SPR) sensor chips by using confined sessile drop technique. A liquid optical adhesive (NOA 61) was dropped on the circular flat surface of cylindrical substrates made of poly(dimethylsiloxane) (PDMS). The formation of hemispherical optical prisms was accomplished by taking advantage of the sharp edges of cylindrical PDMS substrates that prevented the overflow of liquid NOA 61 at the edge of substrates. The size of the hemispherical optical prisms can be controlled by changing the diameter of the cylindrical PDMS substrates. After UV curing, the SPR sensor chips were obtained by the deposition of 3 nm thick chromium and 47 nm thick gold on the flat side of the prisms. The fabricated miniature SPR sensor chips were then mounted on a three-dimensional-printed flow cell to complete the microfluidic SPR sensor module. The miniature SPR sensor chips provided a comparable sensitivity to the conventional high-refractive-index glass SPR chips. To demonstrate the detection capability of nanometer-sized materials, we applied the miniature microfluidic SPR system for monitoring the deposition of layer-by-layer ultrathin films of poly(diallyldimethylammonium chloride)/poly(sodium 4styrenesulfonate) and for detecting human immunoglobulin G.