Multi-Energy and Fast-Convergence Iterative Reconstruction Algorithm for Organic Material Identification Using X-ray Computed Tomography.

In order to significantly reduce the computing time while, at the same time, keeping the accuracy and precision when determining the local values of the density and effective atomic number necessary for identifying various organic material, including explosives and narcotics, a specialized multi-stage procedure based on a multi-energy computed tomography investigation within the 20-160 keV domain was elaborated. It consisted of a compensation for beam hardening and other non-linear effects that affect the energy dependency of the linear attenuation coefficient (LAC) in the chosen energy domain, followed by a 3D fast reconstruction algorithm capable of reconstructing the local LAC values for 64 energy values from 19.8 to 158.4 keV, and, finally, the creation of a set of algorithms permitting the simultaneous determination of the density and effective atomic number of the investigated materials. This enabled determining both the density and effective atomic number of complex objects in approximately 24 s, with an accuracy and precision of less than 3%, which is a significantly better performance with respect to the reported literature values.

Force Mapping Study of Actinoporin Effect in Membranes Presenting Phase Domains.

Equinatoxin II (EqtII) and Fragaceatoxin C (FraC) are pore-forming toxins (PFTs) from the actinoporin family that have enhanced membrane affinity in the presence of sphingomyelin (SM) and phase coexistence in the membrane. However, little is known about the effect of these proteins on the nanoscopic properties of membrane domains. Here, we used combined confocal microscopy and force mapping by atomic force microscopy to study the effect of EqtII and FraC on the organization of phase-separated phosphatidylcholine/SM/cholesterol membranes. To this aim, we developed a fast, high-throughput processing tool to correlate structural and nano-mechanical information from force mapping. We found that both proteins changed the lipid domain shape. Strikingly, they induced a reduction in the domain area and circularity, suggesting a decrease in the line tension due to a lipid phase height mismatch, which correlated with proteins binding to the domain interfaces. Moreover, force mapping suggested that the proteins affected the mechanical properties at the edge, but not in the bulk, of the domains. This effect could not be revealed by ensemble force spectroscopy measurements supporting the suitability of force mapping to study local membrane topographical and mechanical alterations by membranotropic proteins.