In the past two decades, microfluidics‐based particle production is widely applied for multiple biological usages. Compared to conventional bulk methods, microfluidic‐assisted particle production shows significant advantages, such as narrower particle size distribution, higher reproducibility, improved encapsulation efficiency, and enhanced scaling‐up potency. Herein, an overview of the recent progress of the microfluidics technology for nano‐, microparticles or droplet fabrication, and their biological applications is provided. For both nano‐, microparticles/droplets, the previously established mechanisms behind particle production via microfluidics and some typical examples during the past five years are discussed. The emerging interdisciplinary technologies based on microfluidics that have produced microparticles or droplets for cellular analysis and artificial cells fabrication are summarized. The potential drawbacks and future perspectives are also briefly discussed.