High Resolution Membrane Structures within Hybrid Lipid‐Polymer Vesicles Revealed by Combining X‐Ray Scattering and Electron Microscopy

Abstract Hybrid vesicles consisting of phospholipids and block‐copolymers are increasingly finding applications in science and technology. Herein, small angle X‐ray scattering (SAXS) and cryo‐electron tomography (cryo‐ET) are used to obtain detailed structural information about hybrid vesicles with different ratios of 1‐palmitoyl‐2‐oleoyl‐sn‐glycero‐3‐phosphocholine (POPC) and poly(1,2‐butadiene‐block‐ethylene oxide) (PBd ₂₂ ‐PEO ₁₄ , M _s = 1800 g mol ⁻¹ ). Using single particle analysis (SPA) the authors are able to further interpret the information gained from SAXS and cryo‐ET experiments, showing that increasing PBd ₂₂ ‐PEO ₁₄ mole fraction increases the membrane thickness from 52 Å for a pure lipid system to 97 Å for pure PBd ₂₂ ‐PEO ₁₄ vesicles. Two vesicle populations with different membrane thicknesses in hybrid vesicle samples are found. As these lipids and polymers are reported to homogeneously mix, bistability is inferred between weak and strong interdigitation regimes of PBd ₂₂ ‐PEO ₁₄ within the hybrid membranes. It is hypothesized that membranes of intermediate structure are not energetically favorable. Therefore, each vesicle exists in one of these two membrane structures, which are assumed to have comparable free energies. The authors conclude that, by combining biophysical methods, accurate determination of the influence of composition on the structural properties of hybrid membranes is achieved, revealing that two distinct membranes structures can coexist in homogeneously mixed lipid‐polymer hybrid vesicles.