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Dual-view OPM (dOPM) is an optically folded form of OPM. We present an improved dOPM system employing a 60\u00d7\/1.2NA water immersion primary objective and measure the spatial resolution and fluorescence collection efficiency for illumination angles of 35\u00b0 and 45\u00b0 with respect to the coverslip. Illumination at 35\u00b0 provides slightly better lateral resolution and collection efficiency. Collection efficiency measurements are compared to a full vectorial raytracing simulation of the system. Using a light-sheet angle of 35\u00b0, the median bead FWHM for 100\u2005nm diameter fluorescent beads in x, y, and z and the optical sectioning strength were measured over a volume of 100 \u00d7 100 \u00d7 100\u2005\u03bcm3 to be 0.29, 0.31, 0.83, and 2.45\u20133.00\u2005\u03bcm, respectively when the two dOPM views are fused. We demonstrate less photobleaching in time-lapse dOPM of live mEmerald-expressing organoids compared to widefield epi-fluorescence z-stack imaging under the condition of equal detected fluorescence signal from a point object in focus. We demonstrate dOPM for multifield-of-view 3D imaging of biological samples in 96-well plates and apply it to imaging cells in collagen gel and quantifying the FUCCI cell-cycle reporter to provide drug dose\u2013response curves in spheroids. We also use it to perform time-lapse multifield-of-view imaging and demonstrate the detection of organoid lumen closure and reopening, organoid migration within a collagen gel and observing dynamic events in arrays of ex vivo tissue slices.<\/jats:p>","DOI":"10.1093\/pnasnexus\/pgaf370","type":"journal-article","created":{"date-parts":[[2025,11,26]],"date-time":"2025-11-26T02:29:57Z","timestamp":1764124197000},"source":"Crossref","is-referenced-by-count":8,"title":["High content 3D imaging by dual-view oblique plane microscopy"],"prefix":"10.1093","volume":"4","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9900-6610","authenticated-orcid":false,"given":"Hugh","family":"Sparks","sequence":"first","affiliation":[{"name":"Light Community, Department of Physics, Imperial College London , South Kensington Campus, London SW7 2AZ ,","place":["United Kingdom"]},{"name":"Tumour Cell Biology Laboratory, The Francis Crick Institute , 1 Midland Road, London NW1 1AT ,","place":["United 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