This article reviews the recent advances in microfluidic-chip integrated optical biosensors for simultaneous detection of multiple analytes. In particular, the principles and recent progress in different kinds of multiplex optical biosensors and their biological application were reviewed comprehensively. Sensors based on multiplexed detection have absolute advantages in analysis throughput than single assay. The microfluidic chip, a type of micro-total analysis system (µTAS), provides an ideal platform for integration of high-throughput biosensors. Compared with electronic biosensors, benefitted from the technical development in Micro-Electro-Mechanical System, there have been greater advances in the fabrication of optical sensors and microfluidic chip, and then promoting microfluidic-chip integrated optical biosensors for simultaneous detection of multiple analytes.