Cell analysis at the single-cell level is of great importance to investigate the inherent heterogeneity of cell populations and to understand the morphology, composition, and function of individual cells. With the continuous innovation of analytical techniques and methods, single-cell analysis on microfluidic chip systems has been extensively applied for its precise single-cell manipulation and sensitive signal response integrated with various detection techniques, such as optical, electrical, and mass spectrometric analyses. In this review, we focus on the specific optical events in single-cell analysis on a microfluidic chip system. First, the four most commonly applied optical technologies, i.e., fluorescence, surface-enhanced Raman spectroscopy, surface plasmon resonance, and interferometry, are briefly introduced. Then, we focus on the recent applications of the abovementioned optical technologies integrated with a microfluidic chip system for single-cell analysis. Finally, future directions of optical technologies for single-cell analysis on microfluidic chip systems are predicted.