Single-cell analysis has revolutionized our understanding of cellular heterogeneity, disease mechanism, and personalized medicine. Traditional bulk assays are bound to mask minute but critical differences between cells, necessitating the need for tools that single out, manipulate, and read out cells with single-cell resolution. Microfluidics is now an unparalleled platform for single-cell research, with unheralded control over the microenvironment of cells, reagent consumption, and high-throughput manipulation. This chapter provides a general overview of microfluidic methods in single-cell manufacturing, encapsulation, isolation, and analysis. Focus is on droplet-based, valve-based, and inertial microfluidic systems, each of which has been optimized to address specific biological and biomedical challenges. Furthermore, we explore the integration of microfluidics with downstream analytical devices such as single-cell RNA sequencing, proteomics, and real-time imaging. We also survey recent advances in on-chip cell culture, viability screening, and intracellular sensing. Throughput, cell viability, and integration with omics technologies are examined critically as the challenges. Finally, we highlight upcoming microfluidics-enabled single-cell analysis trends, including organ-on-chip platforms, artificial intelligence collaboration, and clinical applications in diagnostics and therapeutics.