Protocol for microfluidic-based high-precision general polarization fluorescence microscopy of lipid packing in membrane vesicles

Authors:

Gaukhar Zhurgenbayeva, Nika Otrin, Chee Seng Man, Christian Eggeling, Ziliang Zhao

Keywords:

lipid packing; membrane vesicles; lipid packing, protocol

Abstract:

Lipid packing plays a critical role in membrane organization and function, but its precise quantification remains challenging in fluidic environments. Here, we present a protocol to integrate confocal fluorescence generalized polarization (GP) imaging with a polydimethylsiloxane (PDMS)-based microfluidic platform. We describe steps for lipid preparation, giant unilamellar vesicle (GUV) fabrication, microfluidic trapping, and GP analysis using polarity-sensitive dyes. This protocol enables robust measurement of membrane order and is readily adaptable to diverse lipid compositions and aqueous environments.

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