{"database":"bioimages","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Hugh Sparks"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-BIAD2314"],"repository":["bioimages"],"figure_sub":["Specimen","Funding","Study Component","Biosample","organisation","Associations","Image acquisition"],"pubmed_authors":["Julien Colombelli","Erik Sahai","Theresa Suckert","Yuriy Alexandrov","Hugh Sparks","Martin Lee","Mar Arias Garcia","Joffrey Pelletier","Shengjie Zhang","Thomas A. Phillips","Neil O. Carragher","Chris Dunsby","Eduard Batlle","Claudia Owczarek","Nils Gustafsson","Leo Rowe-Brown","Nathan Curry","Wenzhi Hong","Zhizhen Xu","Alix Le Marois","Colin D. H. Ratcliffe","Edwin Garcia","Montserrat Llanses","Chris Bakal","Maddy Parsons","Liuba Dvinskikh","Jayne Culley","Giorgio Stassi","Carme Cortina"],"additional_accession":[]},"is_claimable":false,"name":"High Content 3D Imaging by Dual-View Oblique Plane Microscopy","description":"Oblique 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.","dates":{"release":"2025-11-30T00:00:00Z","modification":"2026-04-23T20:39:57.84Z","creation":"2025-09-25T19:28:14.605Z"},"accession":"S-BIAD2314","cross_references":{}}