Strain localization, a cross-scale issue degrading homogeneity and formability during hot working of TA15 titanium alloy, is complicated by lamellar microstructure for hierarchical feature, spatial morphology, and orientation relationship. In this work, by taking advantage of hat-shaped specimens to maintain deformation mode, the effect of heterogeneous lamellar microstructure on strain localization is accessible and visualized by cross-section comparison enlightened by cake cutting. Moreover, the evolution behavior and mechanism are also investigated by the two-scale crystal plasticity model. The mechanism of strain localization during hot working of lamellar microstructure is re-recognized as the structural softening governed by an abrupt phase interface weakening and continuous geometrical softening. The driving force of strain localization is the ‘collapse’ of the stress gradient between constituent phases intensified by the progression of compelled uniformed deformation. The key to microstructure-dependent strain localization is the influence of lamellar orientation on germination and colony interface on intensification. Colonies with lamellar orientation deviated 45 o from the direction of maximum shear stress are endowed with the intrinsic potential of strain localization. Large-size and strongly-orientated colony interfaces intensify strain localization. These findings provide new insights into microstructure-dependent strain localization supporting a more efficient design of processing routing to control localization powerfully.