This dataset includes sharp-blur pairs of Leishmania image, which is a protozoan parasite microscopy image dataset of Leishmania, obtained from the preserved slides stained with Giemsa. The paired blur-sharp images are acquired by employing a bright-field microscope (Olympus IX53) with 100× magnification oil immersion objectives.We first capture the sharp images as ground truth, then acquire its corresponding out-of-focus images. The extent and nature of defocusing are random along the optical axis, where the degree of out-of-focus is inconsistent from image-to-image. This dataset includes 764 in-focus and 764 corresponding out-of-focus images, where each image is composed of 2304 × 1728 pixels in 24-bit JPG format.