Quantitative optical microscopy and micromanipulation studies on the lipid bilayer membranes of giant unilamellar...

This manuscript discusses basic methodological aspects of optical microscopy and micromanipulation methods to study membranes and reviews methods to generate giant unilamellar vesicles (GUVs). In particular, we focus on the use of fluorescence microscopy and micropipet manipulation techniques to study composition-structure-property materials relationships of free-standing lipid bilayer membranes. Because their size (∼5-100 μm diameter) that is well above the resolution limit of regular light microscopes, GUVs are suitable membrane models for optical microscopy and micromanipulation experimentation. For instance, using different fluorescent reporters, fluorescence microscopy allows strategies to study membrane lateral structure/dynamics at the level of single vesicles of diverse compositions. The micropipet manipulation technique on the other hand, uses Hoffman modulation contrast microscopy and allows studies on the mechanical, thermal, molecular exchange and adhesive-interactive properties of compositionally different membranes under controlled environmental conditions. The goal of this review is to (i) provide a historical perspective for both techniques; (ii) present and discuss some of their most important contributions to our understanding of lipid bilayer membranes; and (iii) outline studies that would utilize both techniques simultaneously on the same vesicle thus bringing the ability to characterize structure and strain responses together with the direct application of well-defined stresses to a single membrane or observe the effects of adhesive spreading. Knowledge gained by these studies has informed several applications of lipid membranes including their use as lung surfactants and drug delivery systems for cancer.