bioimagesUnknownHugh Sparkshttps://www.ebi.ac.uk/biostudies/studies/S-BIAD2314bioimagesSpecimenFundingStudy ComponentBiosampleorganisationAssociationsImage acquisitionJulien ColombelliErik SahaiTheresa SuckertYuriy AlexandrovHugh SparksMartin LeeMar Arias GarciaJoffrey PelletierShengjie ZhangThomas A. PhillipsNeil O. CarragherChris DunsbyEduard BatlleClaudia OwczarekNils GustafssonLeo Rowe-BrownNathan CurryWenzhi HongZhizhen XuAlix Le MaroisColin D. H. RatcliffeEdwin GarciaMontserrat LlansesChris BakalMaddy ParsonsLiuba DvinskikhJayne CulleyGiorgio StassiCarme CortinafalseHigh Content 3D Imaging by Dual-View Oblique Plane MicroscopyOblique plane microscopy (OPM) is a form of light-sheet fluorescence microscopy (LSFM) employing a single microscope objective at the sample for both fluorescence excitation and detection. Dual-view OPM (dOPM) is an optically folded form of OPM. We present an improved dOPM system employing a 60×/1.2NA water immersion primary objective and measure the spatial resolution and fluorescence collection efficiency for illumination angles of 35° and 45° with respect to the coverslip. Illumination at 35° 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°, the median bead FWHM for 100 nm diameter fluorescent beads in x, y and z and the optical sectioning strength were measured over a volume of 100×100×100 μm³, to be 0.29, 0.31, 0.83 and 2.45-3.00 μm 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 multi-field-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-response curves in spheroids. We also use it to perform time-lapse multi-field-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.2025-11-30T00:00:00Z2026-04-23T20:39:57.84Z2025-09-25T19:28:14.605ZS-BIAD2314