Orius sc1000 11mpx

Transmission electron microscopy (TEM) experiments were performed with a Hitachi H7650 microscope (Hitachi Ltd., Chiyoda, Tokyo, Japan) linked to an ORIUS SC1000 11MPX (GATAN, Pleasanton, USA) camera run by Digital Micrograph software (GATAN)). Material ink samples (10 µL) were placed on carbon-coated copper grids at room temperature. The excess was removed, and the samples were dried for 10 minutes. Before TEM imaging, grids were negatively stained using Uranyl-Less solution (Delta Microscopies, Mauressac France).

The physical characteristics of the NE formulations were assessed by DLS, Zeta potential measurement, and TEM. The hydrodynamic size was determined using a DLS device (Zetasizer Nano ZS; Malvern Instruments, Malvern, UK) and with the NEs diluted to 1:1000 (v/v) (mean of 3 independent measurements performed at 25 °C). The Zeta potential was measured using a Zetasizer Nano ZS device coupled to a Folded Capillary Cell (DTS1060) from Malvern Instruments. Oily droplets were quantified by NTA using a NanoSight NS300 instrument (Malvern Instruments). A Hitachi H7650 transmission electron microscope linked to an ORIUS SC1000 11MPX (Gatan Inc., Pleasanton, CA, USA) camera run by Digital Micrograph (Gatan Inc.) was used to study the NE samples (1:50 dilution, v/v) transferred to a carbon-coated copper grid. Iron concentration was quantified by UV spectrometry as previously described [66 (link)].

Granulometric profile of NPs and SLNs were acquired by Dynamic Light Scattering (DLS), using Malvern Instruments (Zetasizer Nano ZS, Palaiseau, France). NPs/SLNs were diluted at 1:1000 (v/v), then the average size and the polydispersity index were obtained in triplicate at 25 °C. To determine the Zeta potential, NPs/SLNs, at the same dilution, were analyzed using a Zetasizer Nano ZS coupled with Folded Capillary Cell (DTS1060). Short-term stability assessment was performed for a period of one month by visually checking the lack of creaming or phase separation and monitoring the size and zeta potential. Transmission electron microscopy was carried out using a Hitachi (Tokyo, Japan) coupled to an ORIUS SC1000 11MPX (Gatan, Pleasanton, CA, USA). All samples were deposited on carbon-coated grids (Delta Microscopies, Mauressac, France) and dried after 3 min of contact. A staining procedure using uranyless (Delta Microscopies, Mauressac, France) or nanotungstene was used.