title: Configurable low-cost plotter device for fabrication of multi-color sub-cellular scale microarrays.
abstract: 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 10(4)-10(5) 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.
authors.affiliation: TU Dortmund, Fakultät für Chemie und Chemische Biologie, Biologisch-Chemische Mikrostrukturtechnik, Otto-Hahn Str. 6, 44227, Dortmund, Germany.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Arrabito
authors.forename: Hendrik
authors.initials: H
authors.lastname: Schroeder
authors.forename: Kathrin
authors.initials: K
authors.lastname: Schröder
authors.forename: Christian
authors.initials: C
authors.lastname: Filips
authors.forename: Ulrich
authors.initials: U
authors.lastname: Marggraf
authors.forename: Christian
authors.initials: C
authors.lastname: Dopp
authors.forename: Muthukumaran
authors.initials: M
authors.lastname: Venkatachalapathy
authors.forename: Leif
authors.initials: L
authors.lastname: Dehmelt
authors.forename: Philippe I H
authors.initials: PI
authors.lastname: Bastiaens
authors.forename: Andreas
authors.initials: A
authors.lastname: Neyer
authors.forename: Christof M
authors.initials: CM
authors.lastname: Niemeyer
mesh_terms: D003365:Costs and Cost Analysis; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D006801:Humans; D061986:MCF-7 Cells; D046210:Microfluidic Analytical Techniques / Q000191:economics / Q000295:instrumentation; D020411:Oligonucleotide Array Sequence Analysis / Q000191:economics / Q000295:instrumentation*; D011327:Printing / Q000295:instrumentation; D059010:Single-Cell Analysis / Q000191:economics / Q000295:instrumentation*; D046888:Tissue Array Analysis / Q000191:economics / Q000295:instrumentation*; D014584:User-Computer Interface
chemical_list: D004815:Epidermal Growth Factor; C512478:EGFR protein, human; D066246:ErbB Receptors
keywords: DNA-directed protein immobilization; EGF receptors; device automation; multiplexed patterns; polymer pen lithography