Generation of primary hepatocyte microarrays by piezoelectric printing

We demonstrate a single-step method for the generation of collagen and poly-l-Lysine (PLL) micropatterns on a poly(ethylene glycol) (PEG) functionalized glass surface for cell based assays. The method involves establishing a reliable silanization method to create an effective non-adhesive PEG layer on glass that inhibits cell attachment, followed by the spotting of collagen or PLL solutions using non-contact piezoelectric printing. We show for the first time that the spotted protein micropatterns remain stable on the PEG surface even after extensive washing, thus significantly simplifying protein pattern formation. We found that adherence and spreading of NIH-3T3 fibroblasts was confined to PLL and collagen areas of the micropatterns. In contrast, primary rat hepatocytes adhered and spread only on collagen micropatterns, where they formed uniform, well defined functionally active cell arrays. The differing affinity of hepatocytes and NIH-3T3 fibroblasts for collagen and PLL patterns was used to develop a simple technique for creating a co-culture of the two cell types. This has the potential to form structured arrays that mimic the in vivo hepatic environment and is easily integrated within a miniaturized analytical platform for developing high throughput toxicity analysis in vitro.

The application of fixed hydrophobic patterns for confinement of aqueous solutions in proteomic microarrays

A protein microarray hybridisation system has been implemented by employing patterned hydrophobic thin films on hydrophilic substrates as a means of confinement for aqueous samples. This approach has the ability to handle, and keep separate, small sample volumes of just a few microlitres. In addition, the system is more straightforward to use than the existing multi-well gasket solution. The paper describes the fabrication method and the system is demonstrated for a model protein microarray assay. © 2011 American Institute of Physics.