Particle size and shape are key properties for the successful utilization of LNPs in various applications. The size and morphology of LNPs are often characterized by a combination of different analytical techniques, including light scattering (i.e., batch DLS) and microscopic methods (e.g., AFM, SEM). Instead of relying on multiple methods, this study aims to provide a comprehensive approach to determine the particle size and shape in a single AF4-MALLS-DLS-RI measurement. To verify the suitability of the developed AF4 method four different LNP samples (i.e., LNP-1, LNP-2, LNP-3 and LNP-4) were analyzed by batch DLS, AFM and SEM for comparison. Moreover, we compared two DLS devices from different manufacturers for LNP analysis, i.e., Zetasizer Nano-ZS (Malvern Panalytical) and DynaPro NanoStar (Wyatt Technologies).