Single-shot high-contrast multiplane imaging with a z-splitter prism

We present a single-shot multiplane widefield imaging strategy using a z-splitter prism, which can be assembled from off-the-self components and only requires a single camera. We further introduce a novel extended-volume 3D deconvolution strategy to suppress far-out-of-focus fluorescence background to significantly improve the contrast of our recorded images, conferring to our system a capacity for quasi optical sectioning. By swapping in different z-splitter configurations, we can prioritize high speed or large 3D field-of-view imaging depending on the application of interest. Moreover, our system can be readily applied to a variety of imaging modalities in addition to fluorescence, such as phase-contrast and darkfield imaging, making it a versatile tool for a wide range of biological or biomedical imaging applications.