Imaging properties of extended depth of field microscopy through single-shot focus scanning

Although the single-shot focus scanning technique (SSFS) has been experimentally demonstrated for extended depth of field (EDOF) imaging, few work has been performed to characterize its imaging properties and limitations. In this paper, based on an analytical model of a SSFS system, we examined the properties of the system response and the restored image quality in relation to the axial position of the object, scan range, and signal-to-noise ratio, and demonstrated the properties via a prototype of 10 × 0.25 NA microscope system. We quantified the full range of the achievable EDOF is equivalent to the focus scan range. We further demonstrated that the restored image quality can be improved by extending the focus scan range by a distance equivalent to twice of the standard DOF. For example, in a focus-scanning microscope with a ± 15 μm standard DOF, a 120 μm focus scan range can obtain a ± 60 μm EDOF, but a 150 μm scan range affords noticeably better EDOF images for the same EDOF range. These results provide guidelines for designing and implementing EDOF systems using SSFS technique.