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Updated: Apr 30, 2026
Single Molecule Fluorescence Microscopy on Planar Supported Bilayers
Published on: October 31, 2015
Protocol for microfluidic-based high-precision general polarization fluorescence microscopy of lipid packing in
Gaukhar Zhurgenbayeva1, Nika Otrin2, Chee Seng Man3
1Institute of Applied Optics and Biophysics, Friedrich-Schiller-Universität Jena, Jena, Germany; Leibniz Institute of Photonic Technologies, Member of the Leibniz Centre for Photonics in Infection Research (LPI), Jena, Germany.
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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. For complete details on the use and execution of this protocol, please refer to Zhao et al.1,2.
