Creating biological membranes on the micron scale: forming patterned lipid bilayers using a polymer lift-off technique.
We present a new method for creating patches of fluid lipid bilayers with conjugated biotin and other compounds down to 1 microm resolution using a photolithographically patterned polymer lift-off technique. The patterns are realized as the polymer is mechanically peeled away in one contiguous piece in solution. The functionality of these surfaces is verified with binding of antibodies and avidin on these uniform micron-scale platforms. The biomaterial patches, measuring 1 micro m-76 microm on edge, provide a synthetic biological substrate for biochemical analysis that is approximately 100x smaller in width than commercial printing technologies. 100 nm unilamellar lipid vesicles spread to form a supported fluid lipid bilayer on oxidized silicon surface as confirmed by fluorescence photobleaching recovery. Fluorescence photobleaching recovery measurements of DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiIC(18)(3))) stained bilayer patches yielded an average diffusion coefficient of 7.54 +/- 1.25 microm(2) s(-1), equal to or slightly faster than typically found in DiI stained cells. This diffusion rate is approximately 3x faster than previous values for bilayers on glass. This method provides a new means to form functionalized fluid lipid bilayers as micron-scale platforms to immobilize biomaterials, capture antibodies and biotinylated reagents from solution, and form antigenic stimuli for cell stimulation.
Department of Biomedical Engineering, School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, USA. rno1@cornell.edu
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Orth
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Kameoka
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Zipfel
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Ilic
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Webb
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Clark
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Craighead
D000327:Adsorption; D001360:Avidin / Q000737:chemistry*; D040761:Biomimetic Materials / Q000138:chemical synthesis / Q000737:chemistry*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D001710:Biotin / Q000737:chemistry*; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis / Q000737:chemistry*; D004800:Enzymes, Immobilized; D036681:Fluorescence Recovery After Photobleaching; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D008051:Lipid Bilayers / Q000138:chemical synthesis / Q000737:chemistry*; D008422:Materials Testing; D008567:Membranes, Artificial; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry; D010781:Photography / Q000379:methods; D012825:Silicon; D013499:Surface Properties
D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D010714:Phosphatidylethanolamines; D010743:Phospholipids; D001360:Avidin; C007510:1,2-dipalmitoyl-3-phosphatidylethanolamine; D001710:Biotin; D012825:Silicon