Lyra 3 scanning electron microscope

The samples were fixed in 2.5% glutaraldehyde for 7 h followed by a process of dehydration through rinsing in an increasing ethanol gradient [40 ]. Modifications of previous protocol were carried out for the final steps. For sample mounting, briefly, a drop of approximately 5 uL of each sample was placed on an aluminum support and dried at room temperature for its posterior SEM observation and metal semi quantitation with energy-dispersive X-ray spectroscopy (EDS) using a JEOL JSM-6490LV (JEOL, Tokyo, Japan) scanning electron microscope equipped with an Oxford INCA PentaFetX3 EDS detector.
Samples were prepared for metal determination inside the cell, using a modification of previous protocols [41 (link)]. After glutaraldehyde fixation, a gradient between ethanol and resin was carried out until the sample was 100% embedded in just resin. Then, the sample was heated for 24 h at 56 °C. After polymerization, samples were cut using Leica ULTRACUT UC7 microtome to obtain 70 nm thick slices, which were placed on 200 mesh cupper grids. Uranyl acetate was used as contrast agent. Samples were observed using a Tescan LYRA 3 Scanning electron microscope (TESCAN, Brno, Czech Republic).

LOM images were taken using microscopes Neophot32 (Karl Zeiss, Jena, Germany), Epiphot (Nikon, Tokyo, Japan) and DSX1000 (Olympus Corp., Tokyo, Japan). SEM was performed using the Jeol JSM-7600F scanning electron microscope (JEOL, Tokyo, Japan) equipped with detectors for EDS (Oxford X-Max 50 mm², Oxford Instruments, Abington, UK) and EBSD (HKL Nordlys, Oxford Instruments, Abington, UK). Quantitative EDS point analysis on the Beraha etched samples was performed with a Tescan Lyra3 scanning electron microscope (Tescan, Brno, Czech Republic). The results of the EBSD analysis in the IPF maps and the grain size map were processed using the ATEX software (Version 3.x, Metz, France) [14 ]. The Bruker D8 Discover powder X-ray diffractometer equipped with Co anode and Lynx Eye detector was used for phase identification and quantification. (Bruker, Billerica, MA, USA)