Jem 2000fx tem

In this study, various characterization methods were used to characterize the samples, including scanning electron microscopy (SEM; FEI Inspect F50, Lausanne, Switzerland), atomic force microscopy (AFM; Park NX 10, Suwon, Korea), Raman spectroscopy (Renishaw, InVia, 532 nm, 100× objective, Wotton-under-Edge, Gloucestershire, England), X-ray photoelectron spectroscopy (XPS; PHI 5000 Versa Probe-II, Lafayette, LA, USA), energy-dispersive spectroscopy (EDS; JEOL JSM-7001 LVF Field Emission SEM), photoluminescent spectroscopy (PL; Nanolog spectrofluorometer, Horiba, Kyoto-shi, Japan), high-resolution transmission electron microscopy (HRTEM; JEOL JEM-2000 FX TEM, 200 kV; JEOL 2100F STEM, 200 kV; Peabody, MA, USA)

The in-situ experiments were performed at the MIAMI-1 facility at the University of Huddersfield³¹ . This consists a low energy, Colutron ion-accelerator coupled to a JEOL JEM-2000FX TEM in which the ions are incident on the specimen at 30° to the direction of the electron beam. The microscope was operated at 80 kV (in order to minimize electron-induced radiation damage) with the specimen horizontal in the TEM (i.e. its surface normal to the electron beam). Implantations were conducted using 3 keV He⁺ ions with a flux of 5 × 10¹⁷ ions/m²/s to a fluence of 2 × 10²¹ ions/m² for implantation at room temperature and with a flux of 4.8 × 10¹⁷ ions/m²/s to a fluence of 3 × 10²¹ ions/m² for an implantation using a liquid-nitrogen-cooled holder at its lowest temperature of 107 K.

Microparticles were fixed in 2.5% glutaraldehyde for 30 min at room temperature, absorbed onto formvar-coated nickel grids recently exposed to glow discharge and negatively stained as described previously (Predescu et al., 2001 (link)). EM grids were analyzed in a JEOL JEM-2000FX TEM. For Alk5 pre-embedding immuno-EM, thick cryostat sections of polyvinylpyrrolidone-fixed tissue were incubated with anti-Alk5 antibody followed by goat anti-rat-IgG conjugated to 8-nm gold and processed by standard EM procedure as described previously (Predescu et al., 1996 (link)).