<p>Cryo-EM analysis was also performed of SORT LNPs with the same JEOL JEM-2200FS microscope at 200kV, allowing visualization of LNPs in their native hydrated state. The images show spherical LNP structures around 100 nm, with some larger aggregates also present (data not shown). These aggregates likely result from interactions between particles due to the non-extrusion-based preparation method, which may explain the variability in particle size. Additionally, particles potentially representing different LNP populations or overlapped structures with low contrast were observed (Figure X). The low sample concentration likely contributed to the limited number of visible particles.</p>
Cryo-EM image of SORT LNPs. The different colored outlines indicate different size populations of LNPs.
</figcaption>
</figure>
</div>
</div>
<p>Overall, while the Cryo-EM data confirm the presence and general morphology of LNPs, variability in size and the presence of aggregates highlight potential areas for optimization, such as refining sample concentration and preparation methods to achieve more consistent particle formation.</p>