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ABSTRACT Enabling efficient injection of light into single-mode fibres (SMFs) is a key requirement in realizing diffraction-limited astronomical spectroscopy on ground-based telescopes. SMF-fed spectrographs, facilitated by the use of adaptive optics (AO), offer distinct advantages over comparable seeing-limited designs, including higher spectral resolution within a compact and stable instrument volume, and a telescope independent spectrograph design. iLocater is an extremely precise radial velocity (EPRV) spectrograph being built for the Large Binocular Telescope (LBT). We have designed and built the front-end fibre injection system, or acquisition camera, for the SX (left) primary mirror of the LBT. The instrument was installed in 2019 and underwent on-sky commissioning and performance assessment. In this paper, we present the instrument requirements, acquisition camera design, as well as results from first-light measurements. Broad-band SMF coupling in excess of 35 per cent (absolute) in the near-infrared (0.97–1.31 ${\mu {\rm m}}$) was achieved across a range of target magnitudes, spectral types, and observing conditions. Successful demonstration of on-sky performance represents both a major milestone in the development of iLocater and in making efficient ground-based SMF-fed astronomical instruments a reality.