Jem 2100f tem system

Immunoelectron microscopy (IEM) was carried out as previously described (Utomo et al. 2020 (link)) with modifications. The purified antigen sample was added to the Cu-grid transmission electron microscopy (TEM) (Nisshin EM Co., Ltd., Tokyo) and incubated for 30 s at room temperature, washed with 30 µl of PBS, and incubated for 30 s, repeated three times. BSA (30 µl of 2% v/v) was used for blocking after adding a purified antigen sample and washed three times with PBS. The Cu-grid was washed in stages. The first and secondary antibodies were anti-E rabbit polyclonal antibody (FUJIFILM Wako Pure Chemical) diluted 30 times, and goat anti-rabbit IgG-conjugated (FUJIFILM Wako Pure Chemical) gold nanoparticles diluted 50 times, respectively. The Cu grid was treated with 1% phosphotungstic acid and analyzed with a JEM-2100F TEM system (JEOL Ltd., Tokyo, Japan).

The cross-sectional morphologies of the films were investigated using a Zeiss AURIGA scanning electron microscope equipped with an energy-dispersive X-ray spectroscopy (EDX) unit in order to evaluate concentration profiles (using built-in sensitivity factors) for Ti and Al across the films’ thicknesses.
In order to obtain complementary information on the microstructural properties of the films, transmission electron microscopy (TEM) was employed. For this purpose, cross-sectional TEM lamellae of samples A and C were machined using a focused ion beam milling workstation (Orsay Physics Cobra Z-05) attached to a Zeiss AURIGA 60 Crossbeam field emission gun scanning electron microscope. The cross-sectional TEM lamellae were glued onto a Cu TEM holder and subsequently polished to a thickness of about 30 nm. The TEM characterization was performed using an image-side-corrected JEOL JEM-2100F TEM system operated at 200 keV in both scanning transmission electron microscopy (STEM) and TEM mode. For the STEM imaging, a 0.7 nm spot size and a high-angle annular dark-field detector were used in order to visualize the Z contrast. The main aim of the TEM analysis was to resolve the nanoscale morphology and microstructure of the self-organized nanolamellae formed in the films.