Abstract
The three-dimensional (3D) images produced by the traditional bright-field (BF) light microscope are ill-defined and therefore require reconstruction. Current alternatives to perform 3D BF imaging require dedicated instrumentation or extensive computer modeling. We report optical sectioning in bright-field microscopy (OSBM), a direct method for 3D imaging of label-free samples, where the BF inverse-imaging problem is solved in real space by using an elemental combination of hardware and software. Our method consists in acquiring z-stack images of minimally treated samples under Köhler illumination in the coherent regime, followed by a digital image processing pipeline designed to localize the source of axial intensity gradients that correspond to the z-locations of scatterers within the sample. We validate OSBM by visualizing fungal, animal tissue, and plant samples and comparing with light sheet fluorescence microscopy imaging, finding excellent match for sparse spatial distributions. Our work demonstrates how the standard microscope can be used as an effective 3D imaging device.
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