The alloy Zr-2.5Nb is used for manufacturing the pressure tubes installed in the core of Canada Deuterium Uranium (CANDU) nuclear reactors. This alloy consists of α-zirconium with hexagonal close-packed (HCP) crystals and a minor β-zirconium phase with body centered cubic (BCC) crystals that are mainly located in between the α-grains. Due to the significant elastic and plastic anisotropy of HCP crystals as well as the development of strong textures during manufacturing, these tubes exhibit significant mechanical anisotropy. However, both local microstructure and texture can change, affecting the local patterning of stress fields and the subsequent hydrogen embrittlement rate. Resolving the local microstructure variation of Zr-2.5Nb is challenging due to the development of nano-sized grains during manufacturing. This paper focuses on developing a procedure for the consistent characterization of such local variations using high spatial resolution electron backscatter diffraction (EBSD) and imaging. Microstructure variations along the radial and transverse directions of the transverse-radial plane, as well as along the axial direction of the transverse-axial and radial-axial planes are studied. It is shown that microstructure variations are non-negligible. For the locations that the transverse-radial planes are studied, pole figures rotate, and Kearns factors decrease along the radial direction. In addition, the morphologies of both α- and β-crystals change significantly along the radial direction. With the procedure used for characterization, some β-crystals are indexed and their orientation relationship with their neighboring α-grains are examined.