{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T05:48:33Z","timestamp":1788500913905,"version":"build-2803163510"},"reference-count":32,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2025,6,12]],"date-time":"2025-06-12T00:00:00Z","timestamp":1749686400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,6,12]],"date-time":"2025-06-12T00:00:00Z","timestamp":1749686400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["513025799"],"award-info":[{"award-number":["513025799"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100019180","name":"HORIZON EUROPE European Research Council","doi-asserted-by":"publisher","award":["850691"],"award-info":[{"award-number":["850691"]}],"id":[{"id":"10.13039\/100019180","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Small Methods"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The development of therapeutics builds on testing their efficiency in vitro. To optimize gene therapies, for example, fluorescent reporters expressed by treated cells are typically utilized as readouts. Traditionally, their global fluorescence signal has been used as an estimate of transduction efficiency. However, analysis in individual cells within a living 3D tissue remains a challenge. Readout on a single\u2010cell level can be realized via fluorescence\u2010based flow cytometry at the cost of tissue dissociation and loss of spatial information. Complementary, spatial information is accessible via immunofluorescence of fixed samples. Both approaches impede time\u2010dependent studies on the delivery of the vector to the cells. Here, quantitative 3D characterization of viral transduction efficiencies in living retinal organoids is introduced. The approach combines quantification of gene delivery efficiency in space and time, leveraging human retinal organoids, engineered adeno\u2010associated virus (AAV) vectors, confocal live imaging, and deep learning\u2010based image segmentation. The integration of these tools in an organoid imaging and analysis pipeline allows quantitative testing of future treatments and other gene delivery methods. It has the potential to guide the development of therapies in biomedical applications.<\/jats:p>","DOI":"10.1002\/smtd.202401050","type":"journal-article","created":{"date-parts":[[2025,6,13]],"date-time":"2025-06-13T01:44:59Z","timestamp":1749779099000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["3D Quantification of Viral Transduction Efficiency in Living Human Retinal Organoids"],"prefix":"10.1002","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-5943-407X","authenticated-orcid":false,"given":"Teresa S.","family":"Rogler","sequence":"first","affiliation":[{"name":"Institute of Biophysics Ulm University  89081 Ulm Germany"},{"name":"Faculty of Physics and Center for NanoScience (CeNS) LMU  80539 Munich Germany"},{"name":"Graduate School of Systemic Neuroscience (GSN)  82152 Munich Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katja 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