Single-cell analysis, including sequencing, imaging, and biochemical assays, has become a fundamental strategy in biomedical research. Microplates, with their open system design, facilitate multistep reagent addition, subtraction, and buffer exchange, while their physically isolated wells prevent cross-contamination between biomolecules, establishing them as foundational compartmentalized platform for single-cell analysis. In contrast, water-in-oil droplets, produced by microfluidic systems, create nanoliter/picoliter-sized droplets that act as advanced compartmentalized platform. Although water-in-oil droplet systems offer significant advantages in single-cell analysis, their nearly complete isolation presents substantial limitations. This isolation impedes the development of ex vivo systems requiring material exchange, complicating complex multistep biochemical reactions and hindering the advancement of single-cell multiomics technologies and nonsequencing applications. Recent innovations in permeability-engineered compartmentalization systems, featuring unique materials and structures with controllable material exchange, promise to overcome these limitations. We discuss the latest advancements in permeability-engineered compartmentalization system, elucidates its underlying principles, and explores its potential applications in the field of single-cell analysis.