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Preparation of Aluminum Oxide-Coated Glass Slides for Glycan Microarrays

机译:用于聚糖微阵列的氧化铝涂层玻片的制备

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In this study, we report the fabrication of aluminum oxide-coated glass (ACG) slides for the preparation of glycan microarrays. Pure aluminum (Al, 300 nm) was coated on glass slides via electron-beam vapor deposition polymerization (VDP), followed by anodization to form a thin layer (50–65 nm) of aluminum oxide (Al-oxide) on the surface. The ACG slides prepared this way provide a smooth surface for arraying sugars covalently via phosphonate formation with controlled density and spatial distance. To evaluate this array system, a mannose derivative of α-5-pentylphosphonic acid was used as a model for the optimization of covalent arraying based on the fluorescence response of the surface mannose interacting with concanavalin A (ConA) tagged with the fluorescence probe A488. The ACG slide was characterized using scanning electron microscopy, atomic force microscopy (AFM), and ellipsometry, and the sugar loading capacity, uniformity, and structural conformation were also characterized using AFM, a GenePix scanner, and a confocal microscope. This study has demonstrated that the glycan array prepared from the ACG slide is more homogeneous with better spatial control compared with the commonly used glycan array prepared from the N -hydroxysuccinimide-activated glass slide.
机译:在这项研究中,我们报告了用于制备聚糖微阵列的氧化铝涂层玻璃(ACG)玻片的制造。通过电子束气相沉积聚合(VDP)将纯铝(Al,300 nm)涂覆在载玻片上,然后进行阳极氧化,在表面上形成氧化铝(Al-oxide)的薄层(50-65 nm)。以此方式制备的ACG载玻片提供了光滑的表面,可通过可控制的密度和空间距离的膦酸酯形成来共价排列糖。为了评估该阵列系统,基于表面甘露糖与用荧光探针A488标记的伴刀豆球蛋白A(ConA)相互作用的荧光响应,将α-5-戊基膦酸的甘露糖衍生物用作优化共价排列的模型。使用扫描电子显微镜,原子力显微镜(AFM)和椭圆光度法对ACG玻片进行表征,并使用AFM,GenePix扫描仪和共聚焦显微镜对糖载量,均匀性和结构构象进行表征。这项研究表明,与由N-羟基琥珀酰亚胺活化的载玻片制备的常用聚糖阵列相比,由ACG载玻片制备的聚糖阵列更均匀,具有更好的空间控制。

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