One of the goals of point-of-care (POC) is a chip-based, miniaturized, portable, self-containing system that allows the assay of proteins, nucleic acids, and cells in complex samples. The integration of nanomaterials and microfluidics can help achieve this goal. This tutorial review outlines the mechanism of assaying biomarkers by gold nanoparticles (AuNPs), and the implementation of AuNPs for microfluidic POC devices. In line with this, we discuss some recent advances in AuNP-coupled microfluidic sensors with enhanced performance. Portable and automated instruments for device operation and signal readout are also included for practical applications of these AuNP-combined microfluidic chips.