We developed microfluidics platform which satisfies high anti-fouling level required for single molecule fluorescence imaging. The device consists of three channels and each channel can capture up to 150 cells. We captured normal PC9 cells or drug treated PC9 cells and immobilized EGFR, a major target for cancer therapy, onto the surface. Using single molecule Co-Immunoprecipitation (smCo-IP) and Single-molecule pull-down (SimPull) assay technique, interactions between EGFR and plc-γ-eGFP, Grb2-eGFP, and P85-eGFP were observed. Also we measured relative EGFR surface density. We expect this assay will provide a robust platform for studying membrane proteins at single cell level.