{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:50:00Z","timestamp":1792788600235,"version":"4.5.0"},"reference-count":40,"publisher":"The Company of Biologists","issue":"23","license":[{"start":{"date-parts":[[2024,12,10]],"date-time":"2024-12-10T00:00:00Z","timestamp":1733788800000},"content-version":"vor","delay-in-days":9,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"BrightFocus Macular Degeneration","award":["M2022002F"],"award-info":[{"award-number":["M2022002F"]}]},{"name":"BrightFocus Macular Degeneration","award":["M2021001F"],"award-info":[{"award-number":["M2021001F"]}]},{"DOI":"10.13039\/501100000190","name":"University of Alberta","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100000190","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"crossref","award":["224586\/Z\/21\/Z"],"award-info":[{"award-number":["224586\/Z\/21\/Z"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000268","name":"Biotechnology and Biological Sciences Research Council","doi-asserted-by":"crossref","award":["BB\/S010386\/1"],"award-info":[{"award-number":["BB\/S010386\/1"]}],"id":[{"id":"10.13039\/501100000268","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000765","name":"University College London","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100000765","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,12,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>To understand the multicellular composition of tissues, and how it is altered during development, ageing and\/or disease, we must visualise the complete cellular landscape. Currently, this is hindered by our limited ability to combine multiple cellular markers. To overcome this, we adapted a highly multiplexed immunofluorescence (IF) technique called \u2018Iterative Bleaching Extends Multiplexity\u2019 (IBEX) to the zebrafish retina. We optimised fluorescent antibody micro-conjugation to perform sequential rounds of labelling on a single tissue to simultaneously visualise all major retinal cell types with 11 cell-specific antibodies. We further adapted IBEX to be compatible with fluorescent transgenic reporter lines, in situ hybridisation chain reaction (HCR), and whole-mount immunofluorescence (WMIF). We applied IBEX at multiple stages to study the spatial and temporal relationships between glia and neurons during retinal development. Finally, we demonstrate the utility of IBEX across species by testing it on the turquoise killifish (Nothobranchius furzeri) and African clawed frog (Xenopus laevis) to glean large amounts of information from precious tissues. These techniques will revolutionise our ability to visualise multiple cell types in any organism where antibodies are readily available.<\/jats:p>","DOI":"10.1242\/jcs.263407","type":"journal-article","created":{"date-parts":[[2024,11,14]],"date-time":"2024-11-14T05:11:53Z","timestamp":1731561113000},"source":"Crossref","is-referenced-by-count":5,"title":["\u2018Iterative Bleaching Extends Multiplexity\u2019 facilitates simultaneous identification of all major retinal cell types"],"prefix":"10.1242","volume":"137","author":[{"given":"Aanandita A.","family":"Kothurkar","sequence":"first","affiliation":[{"name":"Institute of Ophthalmology, University College London 1 , London EC1V 9EL ,","place":["UK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gregory S.","family":"Patient","sequence":"additional","affiliation":[{"name":"Institute of Ophthalmology, University College London 1 , London EC1V 9EL ,","place":["UK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9094-6745","authenticated-orcid":false,"given":"Nicole C. L.","family":"Noel","sequence":"additional","affiliation":[{"name":"Institute of Ophthalmology, University College London 1 , London EC1V 9EL ,","place":["UK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aleksandra M.","family":"Krzywa\u0144ska","sequence":"additional","affiliation":[{"name":"Institute of Ophthalmology, University College London 1 , London EC1V 9EL ,","place":["UK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9070-1243","authenticated-orcid":false,"given":"Brittany J.","family":"Carr","sequence":"additional","affiliation":[{"name":"University of Alberta 2 Department of Ophthalmology & Visual Sciences , , Edmonton, AB T5H 3V9 ,","place":["Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2088-8310","authenticated-orcid":false,"given":"Colin J.","family":"Chu","sequence":"additional","affiliation":[{"name":"Institute of Ophthalmology, University College London 1 , London EC1V 9EL ,","place":["UK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4194-8925","authenticated-orcid":false,"given":"Ryan B.","family":"MacDonald","sequence":"additional","affiliation":[{"name":"Institute of Ophthalmology, University College London 1 , London EC1V 9EL ,","place":["UK"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[2024,12,10]]},"reference":[{"key":"2024121012402838300_JCS263407C1","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1146\/annurev.cellbio.042308.113259","article-title":"From progenitors to differentiated cells in the vertebrate retina","volume":"25","author":"Agathocleous","year":"2009","journal-title":"Annu. 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