The Hidden Cost of Two-Dimensional TEM Reconstruction of Magnetic Nanoparticle Morphology
Conventional transmission electron microscopy (TEM) records two-dimensional (2D) projections of three-dimensional (3D) objects, complicating the inference of nanoparticle morphology. This limitation is investigated for quasi-spherical iron oxide nanoparticles (IONPs) imaged after deposition on a TEM grid and after cellular internalization and immobilization in resin. A statistical inverse-reconstruction method is developed in which synthetic 3D particle populations are randomly oriented, projected onto a TEM-like plane, and compared with the experimental equivalent-diameter and axial-ratio distributions. Models of increasing geometrical complexity are considered, from monodisperse prolate spheroids to size- and shape-polydisperse prolate populations and triaxial ellipsoids. Size polydispersity is required to reproduce the projected equivalent-diameter distribution, whereas relaxing axial symmetry through a triaxial description enables simultaneous reproduction of the size and axial-ratio distributions. The inferred population represents a statistically compatible 3D morphology rather than a unique particle-by-particle reconstruction. Its physical relevance is assessed through a macrospin model incorporating the corresponding shape-anisotropy contributions. Application to magnetic fluid hyperthermia reveals a pronounced sensitivity of the predicted heating response to particle geometry and polydispersity. The approach provides a general framework for connecting conventional TEM measurements with morphology-dependent properties of nanoparticle ensembles.