Responsive photonic crystals (PCs) have attracted much attention due to their broad applications in the field of chemical and physical sensing through varying optical properties when exposed to external stimuli. In particular, assembly of block copolymers (BCPs) has proven to be a robust platform for constructing PCs in the form of films or bulk. Here, the generation of BCPs photonic microspheres is presented with 3D periodical concentric lamellar structures through confined self‐assembly. The structural color of the spherical PCs can be tuned by selective swelling of one block, yielding large change of optical property through varying both layer thickness and refraction index of the domains. The as‐formed spherical PCs demonstrate large reflection wavelength shift (≈400–700 nm) under organic solvent permeation and pH adjustment. Spherical shape and structural symmetry endow the formed spherical PCs with rotation independence and monochrome, which is potentially useful in the fields of displays, sensing, and diagnostics. Block copolymer photonic microspheres with onion‐like structure are generated via confined assembly. The structural color of the responsive photonic crystals (PCs) can be tuned by selective swelling, yielding large optical tunability by varying layer thickness and refraction index of the domains. The formed spherical PCs demonstrate a large reflection wavelength shift under solvent permeation and pH adjustment, as well as rotation independence. They are potentially useful in displays, sensing, and diagnostics.