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Nanoscale Patterning of Antigen on Silicon Substrate to Examine Mast Cell Activation

Published online by Cambridge University Press:  01 February 2011

Reid N. Orth
Affiliation:
School of Applied and Engineering Physics
Min Wu
Affiliation:
Department of Chemistry and Chemical Biology, Cornell University
Theodore G. Clark
Affiliation:
Department of Microbiology and Immunology Ithaca, NY 14853
David A. Holowka
Affiliation:
Department of Chemistry and Chemical Biology, Cornell University
Barbara A. Baird
Affiliation:
Department of Chemistry and Chemical Biology, Cornell University
Harold G. Craighead
Affiliation:
School of Applied and Engineering Physics
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Abstract

Rat Basophilic Leukemia (RBL) cells are immobilized and stimulated on micro- and nanometer scale patterns of supported lipid bilayers. The patterns are realized as the photolithographically patterned polymer is mechanically peeled away in one contiguous piece in solution. The 0.36 μm2 to 4, 489 μm2 patches can contain both fluorescent lipids and lipid-linked antigen and provide a synthetic biological substrate for analysis of cell surface receptor-mediated events. 100-nm unilamellar lipid vesicles spread to form a supported lipid bilayer on a thermally oxidized silicon surface as confirmed by fluorescence recovery after photobleaching (FRAP). Aggregation of fluorescently labeled receptors is observed as their coincidence with the patterned antigen. Cell morphology is analyzed with scanning electron microscopy (SEM). Thus, a novel method has been developed for patterning antigen, capturing and immobilizing cells via specific receptors, and spatially controlling antigenic stimulus on the nanoscale.

Type
Research Article
Copyright
Copyright © Materials Research Society 2002

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