Photolithographic patterning of organosilane monolayer for generating large area two-dimensional B lymphocyte arrays.
Photolithographic patterning of organosilane monolayer for generating large area two-dimensional B lymphocyte arrays.
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DOI：
10.1039/b810329a
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发表时间：
2008-12
期刊：
影响因子：
6.1
通讯作者：
Ho CM
中科院分区：
文献类型：
--
作者：
Li N;Ho CM
高密度活细胞阵列是高通量免疫分型系统和基于细胞的生物传感器开发的重要工具。在本文中，我们首次展示了一种简单的制造工艺，以形成六甲基二硅氮烷（HMDS）和聚（乙二醇）（PEG）的二元分子表面，可用于有效地形成高保真的细胞阵列。HMDS自组装单分子膜（SAM）在玻璃基底上的图形化和其物理吸附蛋白质的能力分别通过接触角测量和荧光显微镜表征。经验证，PEG对非HMDS包被背景的钝化对预图案化HMDS没有影响，并极大地抑制了非特异性蛋白结合。以生物素-链霉亲和素复合物为中间体，固定化的B淋巴细胞特异性抗CD 19抗体具有良好的定向性和生物活性，从而保证了高分辨率B淋巴细胞阵列的形成。细胞-配体相互作用的特异性进行了研究，抗-CD 19修饰的微图案呈现出更高的细胞捕获率（88%）比那些由非特异性配体修饰的（15%的抗-CD 5和7%的链霉亲和素）。该方法被证实是生物相容性和抗体修饰的表面的性质保持12小时的细胞培养后。HMDS单层形成和图案化过程，以及通用HMDS/生物素-BSA/链霉亲和素模板，提供了一种非常简单和方便的方法来产生细胞粘附配体的高分辨率微图案，并且也可扩展以形成其他类型细胞的阵列。
High-density live cell array serves as a valuable tool for the development of high-throughput immunophenotyping systems and cell-based biosensors. In this paper, we have, for the first time, demonstrated a simple fabrication process to form the hexamethyldisilazane (HMDS) and poly(ethylene glycol) (PEG) binary molecular surface which can be used to effectively form high fidelity cell arrays. The HMDS self-assembled monolayer (SAM) on glass substrates was photolithographically patterned and its ability to physically adsorb proteins was characterized by contact angle measurement and fluorescence microscopy respectively. Passivation of the non-HMDS coated background by PEG was verified to have no impact on the pre-patterned HMDS and greatly inhibited the non-specific protein binding. Using the biotin–streptavidin complexation as an intermediate, uniform orientation and high bioactivity were achieved for the immobilized B lymphocyte specific anti-CD19 antibodies and therefore ensured the formation of high resolution B lymphocyte arrays. The cell–ligand interaction specificity was investigated and the anti-CD19 decorated micropatterns presented a much higher cell-capturing rate (88%) than those modified by non-specific ligands (15% for anti-CD5 and 7% for streptavidin). The approach was verified to be biocompatible and the properties of the antibody-modified surface were maintained after 12 h cell culture. The HMDS monolayer formation and patterning processes, and the universal HMDS/biotin-BSA/streptavidin template, provide a very simple and convenient process to generate high resolution micropatterns of cell-adhesive ligands and are extendable to form arrays of other types of cells as well.
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影响因子：
3
作者：
HIROTSU, T
通讯作者：
HIROTSU, T
影响因子：
6.2
作者：
Kim, H;Doh, J;Hammond, PT
通讯作者：
Hammond, PT
影响因子：
12.6
作者：
Lan, S;Veiseh, M;Zhang, MQ
通讯作者：
Zhang, MQ
影响因子：
2.1
作者：
Luo, Jen-Tsung;Wu, Wen-Fa;Chen, Wu-Nan
通讯作者：
Chen, Wu-Nan
影响因子：
3.9
作者：
Pardo, L;Wilson, WC;Boland, TJ
通讯作者：
Boland, TJ