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• W2019098052 abstract "Purpose In eye banks, corneal quality control requires an accurate endothelial cell density (ECD). ECs are counted after osmotic preparation (NaCl/sucrose) that makes cells visible. Nevertheless, endothelial images remain often poorly contrasted, non uniform, with a high background noise. Aim: to replace this technique by a non toxic ‘endothelial dye’Methods 30 dyes were pre-selected. Ability to stain EC was assessed on porcine, bovine and rabbit corneas. They were exposed (1 min) to pure or diluted dyes and observed under a light microscope using bright field and 4 standard fluorescent filters. Toxicity was assessed in vitro on EC cultures (live/dead assay, Hoechst Ethidium Calcein). Using the same assay, the less toxic dyes were further assessed on keratocytes cultures and on animal corneas organ cultured for 24H after exposure to dyes. Controls: exposure to BSS only or 3% H2O2Results Only Ledermycin, Hemalun, Rose Bengal, Nuclear Red, Congo red, Evans Blue, and Chicago Sky Blue (CSB) stained ECs, and only the 3 diazoic dyes (Congo red, Evans Blue, and CSB) stained EC borders using fluorescence (Em554/Ex568nm). Both blue dyes also stained the nuclei of dead cells. Hemalun and Rose Bengal were highly toxic. In vitro and ex vivo toxicity of CSB was negligible. The other dyes had an intermediate toxicityConclusion Diazoic dyes and especially CSB could be non toxic dyes for ECs borders. They could improve the accuracy of ECD determination. Their ability to stain human endothelium and their safety (using an animal graft model) have to be assessed as well as their contribution in ECD determination versus the classical osmotic endothelial preparation" @default.
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• W2019098052 date "2008-09-01" @default.
• W2019098052 modified "2023-09-27" @default.
• W2019098052 title "Selection and assessment of vital dyes to improve the endothelial quality control of organ cultured corneas" @default.
• W2019098052 doi "https://doi.org/10.1111/j.1755-3768.2008.5423.x-i1" @default.
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