Abstract：
OBJECTIVE: To evaluate the use of vital dyes and light microscopy for assessing canine corneal endothelial morphology ex vivo.
METHODS: The corneas of 40 canine eyes (n = 20 dogs) enucleated <24 h following euthanasia or death were isolated and flat-mounted on a slide. Corneal endothelium was stained via 0.25% trypan blue followed by 0.5% alizarin red (pH 4.2), photographed, then the following morphological features were calculated using ImageJ: mean cell density (MCD), mean cell area (MCA), polymegathism (coefficient of variation of cell area), and pleomorphism (% hexagonality).
RESULTS: Mean ± standard deviation (range) outcomes were: MCD, 2544 ± 541 cells/mm2 (1750-3922 cells/mm2); MCA, 431 ± 97 μm2 (251-626 μm2); polymegathism, 17 ± 2% (14%-22%); pleomorphism, 84 ± 3% (80%-90%). No significant differences (p ≥ .122) were noted for any outcome between male versus female or brachycephalic versus non-brachycephalic dogs. Young dogs (<10 years) had lower MCA (p = .044), lower pleomorphism (p = .003), and higher MCD (p = .035) when compared to older dogs (≥10 years). Age was significantly (p ≤ .049) correlated with MCA (r = 0.467), MCD (r = -0.476), polymegathism (r = 0.444), and pleomorphism (r = 0.609).
CONCLUSIONS: The combination of vital dyes and light microscopy allowed for clear visualization and evaluation of the corneal endothelium in canine eyes ex vivo. Our findings can be used in future studies to deepen our understanding of the corneal endothelium in health and disease.
METHODS: The corneas of 40 canine eyes (n = 20 dogs) enucleated <24 h following euthanasia or death were isolated and flat-mounted on a slide. Corneal endothelium was stained via 0.25% trypan blue followed by 0.5% alizarin red (pH 4.2), photographed, then the following morphological features were calculated using ImageJ: mean cell density (MCD), mean cell area (MCA), polymegathism (coefficient of variation of cell area), and pleomorphism (% hexagonality).
RESULTS: Mean ± standard deviation (range) outcomes were: MCD, 2544 ± 541 cells/mm2 (1750-3922 cells/mm2); MCA, 431 ± 97 μm2 (251-626 μm2); polymegathism, 17 ± 2% (14%-22%); pleomorphism, 84 ± 3% (80%-90%). No significant differences (p ≥ .122) were noted for any outcome between male versus female or brachycephalic versus non-brachycephalic dogs. Young dogs (<10 years) had lower MCA (p = .044), lower pleomorphism (p = .003), and higher MCD (p = .035) when compared to older dogs (≥10 years). Age was significantly (p ≤ .049) correlated with MCA (r = 0.467), MCD (r = -0.476), polymegathism (r = 0.444), and pleomorphism (r = 0.609).
CONCLUSIONS: The combination of vital dyes and light microscopy allowed for clear visualization and evaluation of the corneal endothelium in canine eyes ex vivo. Our findings can be used in future studies to deepen our understanding of the corneal endothelium in health and disease.
摘要：
目的：评估活体染料和光学显微镜在离体评估犬角膜内皮形态中的应用。
方法:在安乐死或死亡后<24小时摘除的40只犬眼(n=20只狗)的角膜被分离并平坦地固定在载玻片上。角膜内皮用0.25%锥虫蓝染色，然后用0.5%茜素红染色（pH4.2），拍照,然后使用ImageJ计算以下形态特征：平均细胞密度（MCD），平均细胞面积（MCA），多聚性(细胞面积的变异系数)，和同构(%六边形)。
结果：平均值±标准偏差（范围）结果为：MCD，2544±541细胞/mm2（1750-3922细胞/mm2）；MCA，431±97μm2(251-626μm2)；聚集派，17±2%(14%-22%)；多态性，84±3%(80%-90%)。男性与女性或短头与非短头犬之间的任何结果均无显着差异（p≥.122）。幼犬（<10岁）的MCA较低（p=.044），较低的多态性（p=0.003），与老年犬（≥10岁）相比，MCD更高（p=.035）。年龄与MCA显著相关(p≤0.049)(r=0.467)，MCD(r=-0.476),polymegathism(r=0.444)，和多态性(r=0.609)。
结论：活体染料和光学显微镜的结合可以清楚地观察和评估犬眼中的角膜内皮。我们的发现可用于未来的研究，以加深我们对健康和疾病中角膜内皮的理解。
方法:在安乐死或死亡后<24小时摘除的40只犬眼(n=20只狗)的角膜被分离并平坦地固定在载玻片上。角膜内皮用0.25%锥虫蓝染色，然后用0.5%茜素红染色（pH4.2），拍照,然后使用ImageJ计算以下形态特征：平均细胞密度（MCD），平均细胞面积（MCA），多聚性(细胞面积的变异系数)，和同构(%六边形)。
结果：平均值±标准偏差（范围）结果为：MCD，2544±541细胞/mm2（1750-3922细胞/mm2）；MCA，431±97μm2(251-626μm2)；聚集派，17±2%(14%-22%)；多态性，84±3%(80%-90%)。男性与女性或短头与非短头犬之间的任何结果均无显着差异（p≥.122）。幼犬（<10岁）的MCA较低（p=.044），较低的多态性（p=0.003），与老年犬（≥10岁）相比，MCD更高（p=.035）。年龄与MCA显著相关(p≤0.049)(r=0.467)，MCD(r=-0.476),polymegathism(r=0.444)，和多态性(r=0.609)。
结论：活体染料和光学显微镜的结合可以清楚地观察和评估犬眼中的角膜内皮。我们的发现可用于未来的研究，以加深我们对健康和疾病中角膜内皮的理解。
