Configurable Low-Cost Plotter Device for Fabrication of Multi-Color Sub-Cellular Scale Microarrays

• Published in: Small (Weinheim an der Bergstrasse, Germany) Germany), 2014-07, Vol.10 (14), p.2870-2876
• Main Authors: Arrabito, Giuseppe, Schroeder, Hendrik, Schröder, Kathrin, Filips, Christian, Marggraf, Ulrich, Dopp, Christian, Venkatachalapathy, Muthukumaran, Dehmelt, Leif, Bastiaens, Philippe I. H., Neyer, Andreas, Niemeyer, Christof M.
• Format: Article
• Language: English
• Subjects: Arrays; Biochips; Construction; Costs and Cost Analysis; Density; Deoxyribonucleic acid; device automation; DNA-directed protein immobilization; EGF receptors; Epidermal Growth Factor - metabolism; Humans; MCF-7 Cells; Microfluidic Analytical Techniques - economics; Microfluidic Analytical Techniques - instrumentation; multiplexed patterns; Multiplexing; Nanotechnology; Oligonucleotide Array Sequence Analysis - economics; Oligonucleotide Array Sequence Analysis - instrumentation; Plotters; polymer pen lithography; Printing; Printing - instrumentation; Receptor, Epidermal Growth Factor - metabolism; Single-Cell Analysis - economics; Single-Cell Analysis - instrumentation; Tissue Array Analysis - economics; Tissue Array Analysis - instrumentation; User-Computer Interface
• ISSN: 1613-6810; 1613-6829
• DOI: 10.1002/smll.201303390

The construction and operation of a low‐cost plotter for fabrication of microarrays for multiplexed single‐cell analyses is reported. The printing head consists of polymeric pyramidal pens mounted on a rotation stage installed on an aluminium frame. This construction enables printing of microarrays onto glass substrates mounted on a tilt stage, controlled by a Lab‐View operated user interface. The plotter can be assembled by typical academic workshops from components of less than 15 000 Euro. The functionality of the instrument is demonstrated by printing DNA microarrays on the area of 0.5 cm2 using up to three different oligonucleotides. Typical feature sizes are 5 μm diameter with a pitch of 15 μm, leading to densities of up to 104–105 spots/mm2. The fabricated DNA microarrays are used to produce sub‐cellular scale arrays of bioactive epidermal growth factor peptides by means of DNA‐directed immobilization. The suitability of these biochips for cell biological studies is demonstrated by specific recruitment, concentration, and activation of EGF receptors within the plasma membrane of adherent living cells. This work illustrates that the presented plotter gives access to bio‐functionalized arrays usable for fundamental research in cell biology, such as the manipulation of signal pathways in living cells at subcellular resolution. The assembly of a low‐cost polymer pen plotter which allows one to readily fabricate biochips with feature sizes in the low micrometer range is described. As an example, DNA oligonucleotide microarrays are produced and serve to generate subcellular size protein biochips for the precise recruitment and activation of EGF receptors within the plasma membrane of living cells.