(A) The light-sheet is launched from a single primary objective at a (B) 30-degree oblique illumination angle, and rapidly scanned in the Y-direction to acquire a 3D volume. (C) At each intermediate position, a two-dimensional plane along the X and S axes is acquired with a scientific CMOS camera. (D) 100 nm fluorescent beads embedded in agarose. Sharp strip of beads reveals the position of the illumination light-sheet beam waist and scan trajectory along the Y-axis. Parallelepiped data geometry results from oblique illumination and computational shearing of the data. (E) FWHM of beads for different illumination light-sheet NAs, as a function of distance from the coverslip. Raw data, no deconvolution. The confocal parameter of the light-sheets projected onto the Z-axis is 3.0, 7.5, and >12 microns, for NAs of 0.34, 0.16, and 0.06, respectively. (F) Lateral and axial resolutions of surface immobilized fluorescent nanospheres and using an excitation NA of 0.16. Raw data, no deconvolution (G) Dimensions of the field of view for the OPM presented here when operated in a laser-scanning format compared to lattice light-sheet microscopy, eSPIM, a microfluidic-based micromirror, and an atomic force microscopy (AFM)-based cantilever. Lateral dimensions of the figure represent the X and Y axes. (H) Representative point-spread functions for surface immobilized fluorescent nanospheres, before and after deconvolution. Scale Bar: 500 nm. (I) Maximum intensity projection of a MV3 cell expressing genetically encoded multimeric nanoparticles. Deconvolved data is shown. Scale Bar: 10 microns. (J) Fourier Ring Correlation analysis of intracellular resolution. Solid lines show mean value, shaded area is the 95% confidence interval.