Direct observation of conformation of a polymeric coating with implications in microarray applications.
Direct observation of conformation of a polymeric coating with implications in microarray applications.
复制标题
直接观察聚合物涂层的构象对微阵列应用的影响。
DOI：
10.1021/ac801954x
复制
发表时间：
2009
影响因子：
7.4
通讯作者：
Unlü,MSelim
中科院分区：
文献类型：
--
作者：
Yalçin,Ayça;Damin,Francesco;Ozkumur,Emre;diCarlo,Gabriele;Goldberg,BennettB;Chiari,Marcella;Unlü,MSelim
研究了三维聚合物涂层（共聚（DMA-NAS-MAPS））的构象以及聚合物涂层表面上 DNA 链的固定和杂交。构象变化，特别是表面吸附聚合物在水合时的膨胀，与该聚合物涂层在 DNA 微阵列应用中的应用一起被量化。荧光标记的短 DNA 链 (23mers) 与吸附聚合物上的官能团共价连接，用作探针来测量聚合物的溶胀。荧光显微镜技术，光谱自干涉荧光显微镜（SSFM），用于直接测量由于膨胀而导致的荧光团轴向位置的变化，精度达到亚纳米级。此外，还研究了单链 DNA (ssDNA) 和双链 DNA (dsDNA) 探针的固定特性，以及 ssDNA 与目标链的杂交。结果表明，离表面越远的 ssDNA 杂交效率越高，这加强了对这种聚合物涂层作为简单但高效且稳健的 DNA 微阵列表面化学的早期分析。
The conformation of a three-dimensional polymeric coating (copoly(DMA-NAS-MAPS)) and immobilization and hybridization of DNA strands on the polymer coated surface are investigated. A conformational change, specifically the swelling of the surface adsorbed polymer upon hydration, is quantified in conjunction with the application of this polymer coating for DNA microarray applications. Fluorescently labeled short DNA strands (23mers) covalently linked to the functional groups on the adsorbed polymer are used as probes to measure the swelling of the polymer. A fluorescence microscopy technique, Spectral Self-Interference Fluorescence Microscopy (SSFM), is utilized to directly measure the change in axial position of fluorophores due to swelling with subnanometer accuracy. Additionally, immobilization characteristics of single stranded DNA (ssDNA) and double stranded DNA (dsDNA) probes, as well as hybridization of ssDNA with target strands have been studied. The results show that ssDNA further away from the surface is hybridized more efficiently, which strengthens the earlier analysis of this polymeric coating as a simple but highly efficient and robust DNA microarray surface chemistry.