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This approach combines confocal microscopy with fluorescence lifetime imaging to assess both morphological and fluorescence lifetime characteristics of the RPE in murine models. Unlike traditional transpupillary methods, our transscleral approach significantly reduces autofluorescence interference from the cornea and lens, providing clearer images and accurate fluorescence lifetime measurements. Using a continuous\u2010wave 405\u2009nm laser, we achieved high\u2010resolution imaging of the RPE without the need for complex pulsed laser setups. Morphological characterization demonstrated that the transscleral approach effectively visualizes hexagonal, mono\u2010 and binucleated RPE cells with clear delineation. Fluorescence lifetime analysis using pulsed 405\u2009nm laser revealed distinct autofluorescent signatures corresponding to putative endogenous fluorophores. Notably, fixed tissue samples showed altered fluorescence lifetimes compared to fresh samples, highlighting the influence of preservation on biochemical assessment. Quantitative fluorescence lifetime data revealed differences between autofluorescent granules and the surrounding cytoplasm, indicating the potential to obtain information about changes in the cellular chemical environment. Our results suggest that transscleral FLIM imaging is a viable tool for non\u2010destructive RPE analysis, offering potential applications in studying retinal diseases.<\/jats:p>","DOI":"10.1002\/jemt.70133","type":"journal-article","created":{"date-parts":[[2026,2,20]],"date-time":"2026-02-20T04:55:52Z","timestamp":1771563352000},"page":"1016-1027","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Innovative Transscleral Approach to Obtain High Quality Non\u2010Destructive Images of the Retinal Pigment Epithelium"],"prefix":"10.1002","volume":"89","author":[{"given":"Lucia","family":"Dominguez","sequence":"first","affiliation":[{"name":"Instituto de Investigaci\u00f3n y Desarrollo en Bioingenier\u00eda y Bioinform\u00e1tica (IBB), Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET)\u2014UNER  Oro Verde Entre R\u00edos Argentina"},{"name":"Laboratorio de Microscopia Aplicada a Estudios Moleculares y Celulares (LAMAE), Facultad de Ingenier\u00eda\u2014Universidad Nacional de Entre R\u00edos (FI\u2010UNER)  Oro Verde Entre R\u00edos Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Etchart","sequence":"additional","affiliation":[{"name":"Instituto de Investigaci\u00f3n y Desarrollo en Bioingenier\u00eda y Bioinform\u00e1tica (IBB), Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET)\u2014UNER  Oro Verde Entre R\u00edos Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodrigo M.","family":"Torres","sequence":"additional","affiliation":[{"name":"ROMAT Creator Center  Colonia Avellaneda Entre R\u00edos Argentina"},{"name":"Consejo Argentino de Oftalmolog\u00eda (Innovaci\u00f3n y Desarrollo)  Ciudad de Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Javier","family":"Adur","sequence":"additional","affiliation":[{"name":"Instituto de 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