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High-cycle fatigue (HCF) is a critical property of metastable β Ti alloys in aerospace applications. In this work, the HCF behavior and corresponding microscale deformation mechanisms of a metastable Ti-5Al-5Mo-5V-1Cr-1Fe (Ti55511) alloy with a basket-weave structure were investigated. HCF and its deformation mechanisms of a Ti55511 alloy were systematically studied in the deformed condition by using a scanning electron microscope (SEM), a transmission electron microscope (TEM), and electron backscatter diffraction (EBSD). It was found that the Ti55511 alloy exhibited an excellent HCF strength (107 cycles, Kt = 1, R = 0.06) of 738 MPa. The fractographic investigation demonstrated that fatigue striations and secondary cracks were the main features in the crack initiation zone. Dislocation analyses indicated that the HCF deformation of the basket-weave microstructure was mainly affected by the dislocation slipping of the primary α (αp) phase. In addition, the dislocation pile-up at the αp/βtrans interface led to crack initiation. EBSD analyses indicated that the prismatic type slip system of the αp phase was preferentially activated during the HCF deformation process of the Ti55511 alloy, followed by the basal type and pyramid type systems. [ABSTRACT FROM AUTHOR]