Home › Cell Biology › Biomembrane Fabrication by the Solvent-assisted Lipid Bilayer (SALB) Method
Cell Biology JoVE (Open Access) Citable · DOI

Biomembrane Fabrication by the Solvent-assisted Lipid Bilayer (SALB) Method

DOI: 10.3791/53073-v
What you'll learn
  • ✓ Form lipid bilayers using SALB method
  • ✓ Create cholesterol-enriched membrane domains
  • ✓ Measure membrane fluidity via FRAP
  • ✓ Quantify cholesterol in supported membranes
Protocol

Biopharma Insights We present an experimental protocol to form a supported lipid bilayer on solid substrates without using lipid vesicles. We demonstrate a one-step method to form a lipid bilayer on silicon dioxide and gold as well as supported membranes with cholesterol-enriched domain for various biological applications.

Difficulty
intermediate
Total time
~3-4 hrs/sample
Biosafety
BSL-1

Steps

1
Form lipid bilayers on solid supports

Apply lipid solution to a hydrophilic solid support and induce bilayer formation via solvent exchange, achieving a supported lipid bilayer on silicon dioxide or gold.

▶ 01:22
2
Assess membrane fluidity with FRAP

Perform a fluorescence recovery after photobleaching assay to measure lateral mobility of lipids in the formed bilayer.

▶ 03:05
3
Quantify cholesterol content in membrane

Run a cholesterol quantification assay to determine the amount of cholesterol present in the cholesterol-enriched supported membranes.

▶ 05:01
4
Analyze planar bilayer fabrication results

Review the outcomes of the SALB method, including bilayer formation and the presence of cholesterol-enriched domains, as demonstrated in the results section.

▶ 07:51
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