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免疫荧光显微镜和激光剥蚀电感耦合等离子体质谱法在眼部组织切片中对角膜硫酸角质素的成像。

Imaging Keratan Sulfate in Ocular Tissue Sections by Immunofluorescence Microscopy and LA-ICP-MS.

机构信息

Institut für Laboratoriumsmedizin, Klinische Chemie und Pathobiochemie, Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin and Berlin Institute of Health, Augustenburger Platz 1, Berlin 13353, Germany.

Freie Universität Berlin, Fachbereich Biologie, Chemie, Pharmazie, Arnimallee 22, Berlin 14195, Germany.

出版信息

ACS Appl Bio Mater. 2022 Feb 21;5(2):853-861. doi: 10.1021/acsabm.1c01240. Epub 2022 Jan 25.

Abstract

Carbohydrate-specific antibodies can serve as valuable tools to monitor alterations in the extracellular matrix resulting from pathologies. Here, the keratan sulfate-specific monoclonal antibody MZ15 was characterized in more detail by immunofluorescence microscopy as well as laser ablation ICP-MS using tissue cryosections and paraffin-embedded samples. Pretreatment with keratanase II prevented staining of samples and therefore demonstrated efficient enzymatic keratan sulfate degradation. Random fluorescent labeling and site-directed introduction of a metal cage into MZ15 were successful and allowed for a highly sensitive detection of the keratan sulfate landscape in the corneal stroma from rats and human tissue.

摘要

糖胺聚糖特异性抗体可作为监测病理导致细胞外基质改变的有价值工具。在此,通过免疫荧光显微镜以及使用组织冷冻切片和石蜡包埋样本的激光烧蚀 ICP-MS,对角蛋白硫酸盐特异性单克隆抗体 MZ15 进行了更详细的表征。用角蛋白酶 II 预处理可防止样品染色,因此证明了角蛋白硫酸盐的有效酶降解。随机荧光标记和在 MZ15 中引入金属笼的定点导入均成功,并允许在大鼠和人组织的角膜基质中高度敏感地检测角蛋白硫酸盐景观。

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