Controlling the internal structure of block copolymer (BCP) particles has a significant influence on its functionalities. Here, a structure‐controlling method is proposed to regulate the internal structure of BCP Janus colloidal particles using different surfactants. Different microphase separation processes take place in two connected halves of the Janus particles. An order‐order transition between gyroid and lamellar phases is observed in polymeric colloids. The epitaxial growth during the structural transformation from gyroid to lamellar phase undergoes a two‐layered rearrangement to accommodate the interdomain spacing mismatch between these two phases. This self‐assembly behavior can be ascribed to the preferential wetting of BCP chains at the interface, which can change the chain conformation of different blocks. The Janus colloidal particles can further experience a reversible phase transition by restructuring the polymer particles under solvent vapor. We anticipate that the new phase behavior found in Janus particles can not only enrich the self‐assembly study of BCPs but also provide opportunities for various applications based on Janus particles with ordered structures.This article is protected by copyright. All rights reserved