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Versatile multiple protein nanopatterning within a microfluidic channel for cell recruitment studiesElectronic supplementary information (ESI) available: Colloidal particle deposition schematic, intensity profile of fluorescence distributions across the channel, the reversible nature of HIS-tag binding, and cell tracks after removal of P-selectin are given as supplementary materials. Also, a video showing cells rolling on the ICAM1/P-selectin nanopattern is available. See DOI: 10.1039/c5lc00916b
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 microch...
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Main Authors: | , , , |
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Format: | Article |
Language: | English |
Online Access: | Get full text |
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Summary: | 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.
A nanopatterned microfluidic channel with variable protein compositions and potential use in leukocyte recruitment studies and cancer cell extravasation models is presented. |
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ISSN: | 1473-0197 1473-0189 |
DOI: | 10.1039/c5lc00916b |