Optical Sectioning Microscopy Through Single-Shot Lightfield Protocol

Abstract

Optical sectioning microscopy is usually performed by means of a scanning, multi-shot procedure in combination with non-uniform illumination. In this paper, we change the paradigm and report a method that is based in the lightfield concept, and that provides optical sectioning for 3D microscopy images after a single-shot capture. To do this we first capture multiple orthographic perspectives of the sample by means of Fourier-domain integral microscopy (FiMic). The second stage of our protocol is the application of a novel refocusing algorithm that is able to produce optical sectioning in real time, and with no resolution worsening, in the case of sparse fluorescent samples. We provide the theoretical derivation of the algorithm, and demonstrate its utility by applying it to simulations and to experimental data.