{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T02:17:03Z","timestamp":1781230623001,"version":"3.54.1"},"reference-count":17,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2022,5,26]],"date-time":"2022-05-26T00:00:00Z","timestamp":1653523200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Agence Nationale de la S\u00e9curit\u00e9 du M\u00e9dicament (ANSM)"},{"name":"BANCO"},{"name":"EFS"},{"name":"Jean Monnet University"},{"name":"Fondation de France Bourse Berthe Fouassier"},{"name":"Fondation Universit\u00e9 Jean Monnet"},{"name":"Agence de la Biom\u00e9decine"},{"name":"Fondation de l\u2019avenir"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>Purpose: Unlike corneas stored in cold storage (CS) which remain transparent and thin, corneas stored in organoculture (OC) cannot be assessed by specular microscopy (SM), because edema and posterior folds occur during storage and prevent from specular reflection. We previously developed an active storage machine (ASM) which restores the intraocular pressure while renewing the storage medium, thus preventing major stromal edema. Its transparent windows allow multimodal corneal imaging in a closed system. Aim: to present SM of corneas stored in this ASM. Methods: Ancillary study of two preclinical studies on corneas stored for one and three months in the ASM. A prototype non-contact SM was developed (CMOS camera, \u00d710 objective, collimated LED source, micrometric stage). Five non-overlapping fields (935 \u00d7 748 \u03bcm) were acquired in exactly the same areas at regular intervals. Image quality was graded according to defined categories (American Cornea Donor Study). The endothelial cell density (ECD) was measured with a center method. Finally, SMECD was also compared to Hoechst-stained cell nuclei count (HoechstECD). Results: The 62 corneas remained thin during storage, allowing SM at all time points without corneal deconditioning. Image quality varied depending on donors and days of control but, overall, in the 1100 images, we observed 55% of excellent and 30% of good quality images. SMECD did not differ from HoechstECD (p = 0.084). Conclusions: The ASM combines the advantages of CS (closed system) and OC (long-term storage). Specular microscopy is possible at any time in the ASM with a large field of view, making endothelial controls easy and safe.<\/jats:p>","DOI":"10.3390\/jcm11113000","type":"journal-article","created":{"date-parts":[[2022,5,27]],"date-time":"2022-05-27T07:05:07Z","timestamp":1653635107000},"page":"3000","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Specular Microscopy of Human Corneas Stored in an Active Storage Machine"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7676-4080","authenticated-orcid":false,"given":"Thibaud","family":"Garcin","sequence":"first","affiliation":[{"name":"Ophthalmology Department, University Hospital, 42000 Saint-Etienne, France"},{"name":"Corneal Graft Biology, Engineering and Imaging Laboratory, BiiGC, EA2521, Federative Institute of Research in Sciences and Health Engineering, Faculty of Medicine, Jean Monnet University, Rue de la Marandi\u00e8re, 42000 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emmanuel","family":"Crouzet","sequence":"additional","affiliation":[{"name":"Corneal Graft Biology, Engineering and Imaging Laboratory, BiiGC, EA2521, Federative Institute of Research in Sciences and Health Engineering, Faculty of Medicine, Jean Monnet University, Rue de la Marandi\u00e8re, 42000 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chantal","family":"Perrache","sequence":"additional","affiliation":[{"name":"Corneal Graft Biology, Engineering and Imaging Laboratory, BiiGC, EA2521, Federative Institute of Research in Sciences and Health Engineering, Faculty of Medicine, Jean Monnet University, Rue de la Marandi\u00e8re, 42000 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1102-920X","authenticated-orcid":false,"given":"Thierry","family":"Lepine","sequence":"additional","affiliation":[{"name":"Corneal Graft Biology, Engineering and Imaging Laboratory, BiiGC, EA2521, Federative Institute of Research in Sciences and Health Engineering, Faculty of Medicine, Jean Monnet University, Rue de la Marandi\u00e8re, 42000 Saint-Etienne, France"},{"name":"Laboratory Hubert Curien, Optics Institute Graduate School, 42000 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philippe","family":"Gain","sequence":"additional","affiliation":[{"name":"Ophthalmology Department, University Hospital, 42000 Saint-Etienne, France"},{"name":"Corneal Graft Biology, Engineering and Imaging Laboratory, BiiGC, EA2521, Federative Institute of Research in Sciences and Health Engineering, Faculty of Medicine, Jean Monnet University, Rue de la Marandi\u00e8re, 42000 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gilles","family":"Thuret","sequence":"additional","affiliation":[{"name":"Ophthalmology Department, University Hospital, 42000 Saint-Etienne, France"},{"name":"Corneal Graft Biology, Engineering and Imaging Laboratory, BiiGC, EA2521, Federative Institute of Research in Sciences and Health Engineering, Faculty of Medicine, Jean Monnet University, Rue de la Marandi\u00e8re, 42000 Saint-Etienne, France"},{"name":"Institut Universitaire de France, Boulevard Saint-Michel, 75005 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,5,26]]},"reference":[{"key":"ref_1","first-page":"2827","article-title":"Cell death during corneal storage at 4 degrees C","volume":"40","author":"Komuro","year":"1999","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_2","first-page":"2887","article-title":"Apoptosis in the endothelium of human corneas for transplantation","volume":"41","author":"Albon","year":"2000","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1136\/bjo.86.3.306","article-title":"Value of two mortality assessment techniques for organ cultured corneal endothelium: Trypan blue versus TUNEL technique","volume":"86","author":"Gain","year":"2002","journal-title":"Br. J. Ophthalmol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1001\/archopht.121.4.442","article-title":"Prospective, randomized clinical and endothelial evaluation of 2 storage times for cornea donor tissue in organ culture at 31 degrees C","volume":"121","author":"Thuret","year":"2003","journal-title":"Arch. Ophthalmol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1725","DOI":"10.1136\/bjophthalmol-2017-310913","article-title":"The existence of dead cells in donor corneal endothelium preserved with storage media","volume":"101","author":"Kitazawa","year":"2017","journal-title":"Br. J. Ophthalmol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3326","DOI":"10.1167\/iovs.02-1255","article-title":"Predicting endothelial cell loss and long-term corneal graft survival","volume":"44","author":"Armitage","year":"2003","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5907","DOI":"10.1167\/iovs.17-22218","article-title":"Storage of Porcine Cornea in an Innovative Bioreactor","volume":"58","author":"Guindolet","year":"2017","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1641","DOI":"10.1111\/ajt.15238","article-title":"Innovative corneal active storage machine for long-term eye banking","volume":"19","author":"Garcin","year":"2019","journal-title":"Am. J. Transplant."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1097\/TP.0000000000003109","article-title":"Three-month Storage of Human Corneas in an Active Storage Machine","volume":"104","author":"Garcin","year":"2020","journal-title":"Transplantation"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"6018","DOI":"10.1167\/iovs.10-6641","article-title":"Pan-corneal endothelial viability assessment: Application to endothelial grafts predissected by eye banks","volume":"52","author":"Pipparelli","year":"2011","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1097\/ICO.0000000000000114","article-title":"CorneaJ: An imageJ Plugin for semi-automated measurement of corneal endothelial cell viability","volume":"33","author":"Bernard","year":"2014","journal-title":"Cornea"},{"key":"ref_12","unstructured":"Takahashi, E.A. (1996). Method for Computing Morphology of Cornea Endothelium Cells. (US5523213A), U.S. Patent."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2062","DOI":"10.1167\/iovs.06-1043","article-title":"Reproducibility of endothelial assessment during corneal organ culture: Comparison of a computer-assisted analyzer with manual methods","volume":"48","author":"Thuret","year":"2007","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1016\/j.ophtha.2004.10.045","article-title":"An evaluation of image quality and accuracy of eye bank measurement of donor cornea endothelial cell density in the Specular Microscopy Ancillary Study","volume":"112","author":"Lass","year":"2005","journal-title":"Ophthalmology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1080\/02713680500536738","article-title":"Specular microscopy ancillary study methods for donor endothelial cell density determination of Cornea Donor Study images","volume":"31","author":"Benetz","year":"2006","journal-title":"Curr. Eye Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1007\/s10561-017-9624-7","article-title":"Considering 3D topography of endothelial folds to improve cell count of organ cultured corneas","volume":"18","author":"Jumelle","year":"2017","journal-title":"Cell Tissue Bank."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1097\/ICO.0000000000001166","article-title":"Rapid Warming of Donor Corneas Is Safe and Improves Specular Image Quality","volume":"36","author":"Tran","year":"2017","journal-title":"Cornea"}],"container-title":["Journal of Clinical Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0383\/11\/11\/3000\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:18:59Z","timestamp":1760138339000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0383\/11\/11\/3000"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,5,26]]},"references-count":17,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2022,6]]}},"alternative-id":["jcm11113000"],"URL":"https:\/\/doi.org\/10.3390\/jcm11113000","relation":{},"ISSN":["2077-0383"],"issn-type":[{"value":"2077-0383","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,5,26]]}}}