High-resolution confocal microscopy at a single-colloid level was combined with testing the mechanical properties of semicrystalline colloidal films using nanoindentation experiments. Using localization and tracking algorithms, the displacements of each individual colloid in the film could be determined, allowing the identification of the dominant microscopic deformation processes. Despite strong attractive forces between the colloids, structural reorganization of the film was ubiquitous. We observed irreversible rearrangements such as dislocation defects and pile-up behavior. In comparison with amorphous systems, reversible deformations of the film were significantly more pronounced.