For monoolein-based cubosomes stabilised by Pluronic F-127, samples were considerably more turbid compared to similarly sized LLCs of other compositions. As is shown in Fig. F, particles adopted a characteristic petal like morphology, with lamellar leaflets encompassing a highly ordered inner core. Several crystallographic planes can be observed in the particle shown, suggesting a partially faceted structure. The section marked in white was subjected to FFT analysis and miller indices were assigned as before, this time for a hexagonal motif suggesting a [111] direction parallel to the optical axis (Fig. ). For monoolein, the triblock stabilising polymer F-127 is thought to be present both in the bicontinuous cubic core and form a sterically stabilising polymeric corona around the particles. A secondary set of FFT points was observed. This proved useful in preliminary phase assignment as the first and second set of points were in a √2: √4 ratio. Fig. H (iFFT) was used to estimate the interplanar distance (d-spacing) and lattice parameter (a): ~105 Å and 148.5 Å respectively. These values were in excellent agreement with those previously reported for similar lipid/polymer ratios. Finally, phytantriol particles doped with tocopherol acetate (10 mol %) were analysed using cryo-EM, a representative example shown in Fig. K. Differentiating between cubosomes and hexosomes via electron microscopy was non-trivial. Both inverse bicontinuous cubic particles seemed to preferentially orientate such that [111] axes were perpendicular to the sample plane, producing hexagonal FFT patterns in each case. It was therefore important to establish that: a) cubic FFT patterns could be found in these two systems, 33 b) a characteristic ''spinning top'' like morphology could be observed in the presumed hexosomal samples, and c) estimated lattice parameters were in agreement with literature values. All three criteria were met, and hexosomal particles with d10 of approximately 42.5 Å were obtained, via FFT and iFFT analysis (Fig. and). A crest can be observed near the apex of a particle marked with a yellow arrow in Fig. K, identical to those described by Dong et. al., who use a similar composition. Various spiral particles resembling snail shells were also observed, reminiscent of toroidal DNA condensates. These were most likely the same ''spinning-top'' structures viewed down another optical axis. In addition to preliminary phase assignment and particle characterisation, cryo-EM was used to measure the percentage of lamellar vesicles (single bilayer spheres) formed alongside LLCs. This has been noted in the past, particularly for methods involving ethanol evaporation. As much as 50 % vesicles can be present in samples composed of phytantriol, described in full by Akhlaghi et. al. Up to 40 cryo-EM micrographs and 200 particles were counted for each composition. There was a clear bias for the formation of LLCs over vesicles based on these results (shown in Fig. and), confirming the appropriateness of this method for future studies highly dependent on particle morphology. <111> <001> <111> To unambiguously assign the phase behaviour of each particulate system, X-ray scattering techniques were employed, diffraction patterns yielding key information on LLC crystallinity. Lipid pastes are typically required for SAXS analysis (very low water content). As a result, MHF samples were concentrated using centrifugal filters as previously described. Samples were analysed before and after concentrating by DLS to assess degree of particle aggregation, though this was not expected to drastically alter scattering.