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Optical sectioning has been widely employed for inhibiting out-of-focus backgrounds in three-dimensional (3D) imaging of biological samples. However, point scanning imaging or multiple acquisitions for wide-field optical sectioning in epi-illumination microscopy remains time-consuming for large-scale imaging. In this paper, we propose a single-scan optical sectioning method based on the hybrid illumination (HiLo) algorithm with a line-illumination strategy. Our method combines HiLo background inhibition with confocal slit detection. It t