The temperature rise due to high current/power operations and stress caused by the expansion or contraction of electrodes by lithium (de)-intercalation is known to be a degradation factor in lithium-ion batteries (LIBs). Therefore, in this study, a new technique is proposed to simultaneously measure the internal temperature and stress in the 18650-type LIB during its operation. The operando measurement involved retaining a constant gage volume using rotating spiral slits, obtaining X-ray diffraction images using a highly sensitive two-dimensional detector, and employing the sin²ψ method to separate the stress and change in temperature. During the charging process, at the 1C current rate, the anode expansion, owing to the lithium intercalation, induced the radial compressive stress in the Cu anode collector. The radial stress changes of the Cu anode collector were -31 MPa (compression) and 44 MPa (tensile) during the 1C charge and discharge processes, respectively. Moreover, the internal temperature, which was higher than the surface temperature, was calculated by considering the radial stress change during the battery operation. During the 4C cycle, the surface and internal temperatures rose by 26 °C and 42 °C, respectively. The results indicate that the internal temperature and stress in the 18650-LIB were successfully measured during battery operation.