Lovchik Robert, von Arx Corinne, Viviani Angelika, Delamarche Emmanuel
IBM Research GmbH, Zurich Research Laboratory, 8803, Rüschlikon, Switzerland.
Anal Bioanal Chem. 2008 Feb;390(3):801-8. doi: 10.1007/s00216-007-1436-3. Epub 2007 Jul 14.
We present a method for the facile arraying of cells on microstructured substrates which should be suitable for cellular assays in autonomous microfluidic capillary systems (CSs). The CSs, which were designed and microfabricated in Si, have various microfluidic functional elements including reaction chambers wherein cellular arrays are located. Two methods for arraying the cells were explored. In the first method, a hydrophobic alkanethiol was microcontact-printed on the bottom surface of a microfluidic reaction chamber. The subsequent adsorption of protein-repellent alkanethiols around the printed areas and the deposition from solution of fibronectin (FN) on the hydrophobic areas resulted in an adhesive pattern for the attachment of living human breast cancer cells. This method was limited by the formation of cellular clusters, which proved difficult to remove selectively. The second method employed a poly(dimethylsiloxane) elastomer having oval recessed microstructures. The selective coating of the inner walls of the ovals with FN and the blocking of the mesas around the ovals with bovine serum albumin (BSA) permitted single or multiple cells to be arrayed depending on the size of the ovals. The possibility of sealing CSs with cells arrayed on poly(dimethylsiloxane) may provide a versatile platform for high-throughput experimentation down to the single-cell level.
我们提出了一种在微结构化基板上轻松排列细胞的方法,该方法适用于自主微流控毛细管系统(CSs)中的细胞分析。这些CSs是在硅中设计和微制造的,具有各种微流控功能元件,包括细胞阵列所在的反应室。我们探索了两种排列细胞的方法。在第一种方法中,将疏水性链烷硫醇微接触印刷在微流控反应室的底表面。随后在印刷区域周围吸附排斥蛋白质的链烷硫醇,并将纤连蛋白(FN)从溶液中沉积在疏水区域上,从而形成用于附着活的人乳腺癌细胞的粘附图案。该方法受到细胞簇形成的限制,事实证明难以选择性地去除细胞簇。第二种方法使用具有椭圆形凹陷微结构的聚二甲基硅氧烷弹性体。用FN选择性地涂覆椭圆形的内壁,并用牛血清白蛋白(BSA)封闭椭圆形周围的台面,从而可以根据椭圆形的大小排列单个或多个细胞。用排列在聚二甲基硅氧烷上的细胞密封CSs的可能性可为单细胞水平的高通量实验提供一个通用平台。