Preparation of SLBs
This protocol is extracted from research article:
Nano-bio-computing lipid nanotablet
Sci Adv, Feb 22, 2019; DOI: 10.1126/sciadv.aau2124

SLBs were prepared on glass substrates within flow chambers via the vesicle fusion method. A flow chamber was made from a top and bottom glass and a Parafilm spacer (4 mm × 50 mm × 200 μm). The working volume of the glass chamber is ~40 μl. The top slide glass (Paul Marienfeld GmbH & Co. KG) with inlet and outlet holes was cleaned by 5-min bath sonication in DI water and 2-min piranha etching in H2SO4/H2O2 (3:1). After each cleaning procedure, glass substrates were rinsed with sufficient amounts of DI water. The cleaned top glass slide was then passivated with bovine serum albumin (BSA) (10 mg/ml) in 150 mM NaCl phosphate-buffered saline (1× PBS) to prevent SLB formation on the upper side of the chamber. The bottom cover glass (Paul Marienfeld GmbH & Co. KG, Germany) was cleaned by 5-min sonication in DI water followed by 2-min piranha etching. A two-ply Parafilm spacer was then placed between the two glass slides and heat-sealed at 100°C. The freshly extruded SUV solution was diluted to 1 mg/ml in 1× PBS solution and sonicated for 15 min. The SLBs were formed by introducing the sonicated vesicle solution into the flow chamber at 30°C. After 60 min, the flow chamber was gently washed with DI water and 1× PBS. Defects in SLBs were then passivated with BSA (100 μg/ml) in 1× PBS for 45 min. Streptavidin (17 nM) in 1× PBS was then injected into the flow chamber to modify the biotinylated DOPE molecules. After 40 min, the flow chamber was washed with 1× PBS twice. The flow chamber with the streptavidin-modified SLB can be stored up to 3 days in a humidified refrigerator at 4°C. Formation of air bubbles inside the chamber should be avoided in all procedures.

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