Multifocus microscopy enables recording of entire volumes in a single camera exposure. In dense samples, multifocus microscopy is severely hampered by background haze. We developed a scalable multifocus method that incorporates optical sectioning and offers improved axial resolution capabilities. In our method, dubbed SOLIS, a dithered oblique light-sheet scans the sample volume during a single exposure, while fluorescence from each illuminated plane in the sample is mapped onto a line on the camera with a multifocus optical element. A synchronized rolling shutter readout realizes optical sectioning. This data set contains example images, volumetric stacks, and timelapse videos that demonstrate the technique on endothelial cells (AF488-phalloidin labelled actin) and uncleared engineered human heart tissue (AF488-TOM20 labelled mitochondria).

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