Drying and Rehydration of DLPC/DSPC Symmetric and Asymmetric Supported Lipid Bilayers: a Combined AFM and Fluorescence Microscopy Study

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Abstract
This work characterizes the impact of lipid symmetry/asymmetry on drying/rehydration reorganization in phase-separated dilauroylphosphatidylcholine (DLPC)/distearoylphosphatidylcholine (DSPC) supported lipid bilayers (SLBs) at the submicron and micron-scale. In addition the prevention of major drying/rehydration reorganization by the use of trehalose is demonstrated. Even though it was found using fluorescence microscopy that micrometer scale structure is preserved in the presence and absence of trehalose upon drying/rehydration, AFM and FRAP experiments successfully revealed major changes in the phase-separated structure such as defects, obstructions, lipid condensation, collapse structures, and complex incomplete DLPC-DSPC mixing/exchange in the absence of trehalose. In the presence of trehalose the membrane preserves its structure at the nanometer scale and mobility. We found that SLBs with asymmetric domain configurations underwent major rearrangements during drying and rehydration, whereas the symmetric domain configuration mainly rearranged during rehydration, that we hypothesize is related to lower transmembrane cohesiveness or lack of anchoring to the substrate in the case of the asymmetric domains.

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