Preparation and imaging of lipid bilayer-coated silica microspheres

This protocol describes the preparation and imaging of lipid bilayer-coated micrometer-scale glass beads. This protocol was used to generate the data presented in Figure 2d-g in the following publication: Bruggeman et al., POLCAM: Instant molecular orientation microscopy for the life sciences. bioRxiv 2023.02.07.527479 (Feb 2023), doi: https://doi.org/10.1101/2023.02.07.527479 The described protocol is based on a previously published protocol that uses lipid extrusion instead of a tip probe sonicator to generate lipid vesicles: Tingting Wu, Jin Lu and Matthew D. Lew. pixOL: pixel-wise dipole-spread function engineering for simultaneously measuring the 3D orientation and 3D localization of dipole-like emitters. bioRxiv (2022). https://doi.org/10.1101/2021.12.30.474544