Dual-view oblique plane microscopy (dOPM)

We present a new folded dual-view oblique plane microscopy (OPM) technique termed dOPM that enables two orthogonal views of the sample to be obtained by translating a pair of tilted mirrors in refocussing space. Using a water immersion 40x 1.15 NA primary objective, deconvolved image volumes of 200 nm beads were measured to have full width at half maxima (FWHM) of 0.35+/-0.04 microns and 0.39+/-0.02 microns laterally and 0.81+/-0.07 microns axially. The laterally integrated z-sectioning value was 1.33+/-0.45 microns using light-sheet FWHM in the frames of the two views of 4.99+/-0.58 microns and 4.89+/-0.63 microns. To qualitatively demonstrate that the system can reduce shadow artefacts while providing a more isotropic resolution, a multi-cellular spheroid approximately 100 microns in diameter was imaged.