We report on a single-shot diffraction imaging methodology using relativistic femtosecond electron pulses generated by a radio-frequency acceleration-based photoemission gun. The electron pulses exhibit excellent characteristics, including a root-mean-square (rms) illumination convergence of 31 ± 2 μrad, a spatial coherence length of 5.6 ± 0.4 nm, and a pulse duration of approximately 100 fs with (6.3 ± 0.6) × 10⁶ electrons per pulse at 3.1 MeV energy. These pulses facilitate high-quality diffraction images of gold single crystals with a single shot. The rms spot width of the diffracted beams was obtained as 0.018 ± 0.001 Å⁻¹, indicating excellent spatial resolution.