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 40× 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 μm and 0.39±0.02 μm laterally and 0.81±0.07 μm axially. The laterally integrated z-sectioning value was 1.33±0.45 μm using light-sheet FWHM in the frames of the two views of 4.99±0.58 μm and 4.89±0.63 μm. To qualitatively demonstrate that the system can reduce shadow artefacts while providing a more isotropic resolution, a multi-cellular spheroid approximately 100 μm in diameter was imaged.