Multifocus microscopy with optical sectioning and high axial resolution

Multifocus microscopy enables recording of entire volumes in a single camera exposure. In dense samples, multifocus is severely hampered by background haze. Here, we introduce scalable method that incorporates optical sectioning and offers improved axial resolution capabilities. our method, dithered oblique light-sheet scans the sample volume during exposure, while fluorescence from each illuminated plane mapped onto line on with element. A synchronized rolling shutter readout realizes sectioning. We describe technique theoretically verify its improvement demonstrate prototype system beam splitter cascade record monolayers endothelial cells at 35 per second. furthermore image uncleared engineered human heart tissue visualize distribution mitochondria high resolution. Our manages to capture sub-diffraction sized mitochondria-derived vesicles up 30 µm deep into tissue.