title: Versatile multiple protein nanopatterning within a microfluidic channel for cell recruitment studies.
abstract: A novel approach combining self-assembly-based colloidal lithography and polydimethylsiloxane (PDMS) micromolding to generate complex protein nanopatterns for studying the mechanisms of leukocyte extravasation within microchannels is presented. Nanostructured surfaces sealed onto PDMS-molded microchannels are chemically functionalized in situ in an all-aqueous process to generate bi-functional chemical nanopatterns. Subsequent co-immobilization with proteins makes use of common non-covalent coupling (e.g. HIS-tags, FC-tags and biotin-tags), giving nanopatterns of arbitrary combinations of oriented, functional proteins. Up to three different proteins were simultaneously co-immobilized into the microchannel with nanoscale precision, demonstrating the complex patterns. As a proof-of-principle, a mimic of an inflamed endothelium was constructed using a macro- and nanoscale pattern of intercellular adhesion molecule 1 (ICAM1) and P-selectin, and the response of leukocytes through live cell imaging was measured. A clear result on the rolling behavior of the cells was observed with rolling limited to areas where ICAM1 and P-selectin are present. This micro/nano-interface will open new doors to investigations of how spatial distributions of proteins control cellular activity.
authors.affiliation: Interdisciplinary Nanoscience Center (iNANO), Århus University, Gustav Wieds Vej 14, 8000 Århus, Denmark.
authors.forename: A S
authors.initials: AS
authors.lastname: Andersen
authors.forename: W F
authors.initials: WF
authors.lastname: Zheng
authors.forename: D S
authors.initials: DS
authors.lastname: Sutherland
authors.forename: X Y
authors.initials: XY
authors.lastname: Jiang
mesh_terms: D062028:Bioprinting / Q000295:instrumentation*; D002460:Cell Line; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004867:Equipment Design; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry*; D018799:Intercellular Adhesion Molecule-1 / Q000737:chemistry*; D036904:Leukocyte Rolling; D007962:Leukocytes / Q000166:cytology*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D049329:Nanostructures / Q000737:chemistry*; D019007:P-Selectin / Q000737:chemistry*
chemical_list: D004129:Dimethylpolysiloxanes; D055767:Immobilized Proteins; D019007:P-Selectin; D018799:Intercellular Adhesion Molecule-1; C013830:baysilon