Creating Biological Membranes on the Micron Scale: Forming Patterned Lipid Bilayers Using a Polymer Lift-Off Technique

• Published in: Biophysical journal 2003-11, Vol.85 (5), p.3066-3073
• Main Authors: Orth, R.N., Kameoka, J., Zipfel, W.R., Ilic, B., Webb, W.W., Clark, T.G., Craighead, H.G.
• Format: Article
• Language: English
• Subjects: Adsorption; Avidin - chemistry; Biomimetic Materials - chemical synthesis; Biomimetic Materials - chemistry; Biosensing Techniques - instrumentation; Biosensing Techniques - methods; Biotin - chemistry; Coated Materials, Biocompatible - chemical synthesis; Coated Materials, Biocompatible - chemistry; Enzymes, Immobilized; Fluorescence Recovery After Photobleaching; Immunoassay - instrumentation; Immunoassay - methods; Lipid Bilayers - chemical synthesis; Lipid Bilayers - chemistry; Lipids; Materials Testing; Membranes; Membranes, Artificial; Molecular biology; Molecules; Nanotechnology - instrumentation; Nanotechnology - methods; Phosphatidylethanolamines - chemistry; Phospholipids - chemistry; Photography - methods; Silicon; Surface Properties
• ISSN: 0006-3495; 1542-0086
• DOI: 10.1016/S0006-3495(03)74725-0

We present a new method for creating patches of fluid lipid bilayers with conjugated biotin and other compounds down to 1 μm 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 μm–76 μm on edge, provide a synthetic biological substrate for biochemical analysis that is ∼100× 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 μm 2 s −1, equal to or slightly faster than typically found in DiI stained cells. This diffusion rate is ∼3× 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.