Titanium alloys are widely employed in aerospace applications due to an outstanding combination of properties. The variety of loading conditions and microstructures encountered in industrial components is urging the development of microstructure sensitive modeling capabilities. In particular, reliable predictions require a good knowledge of operating deformation mechanisms. The present study aims at providing a thorough characterization of deformation mechanisms in the near-α Ti-6Al-2Sn-4Zr-2Mo alloy using experiments and simulations. The sequential activation of deformation mechanisms in the α phase was monitored in situ during a tensile test carried out in a scanning electron microscope using a combination of slip traces analysis and electron back-scattered diffraction. Basal slip is activated first, and prismatic slip activity, which proceeds at a higher macroscopic stress level, is needed for a significant creep/relaxation to occur. While little evidence of 〈a〉-type pyramidal slip was found, 〈c + a〉 pyramidal slip involved first-order pyramidal planes and operates at stress levels near or above the 0.2% proof stress. Atomic force microscopy characterization of the features of the different slip modes revealed that 〈a〉 slip is coarser and wavier than 〈c + a〉 slip. Twinning, which is usually neglected in such alloys within this strain regime, was observed to be slip stimulated at a plastic strain as low as 0.5%. Crystal plasticity parameters leading to an accurate simulation of the activation sequence of deformation mechanisms were then determined. For this purpose, critical resolved shear stress values were derived for the different slip and twinning modes using specific approaches and subsequently validated using crystal plasticity simulations based on fast-Fourier transforms.