Veleta g2 camera

The complete method used for immune electron microscopy is described by Horvatits et al. [24 (link)]. Briefly, 10 µL of each of the samples was adsorbed onto carbon/formvar -oated copper grids (Plano GmbH, Wetzlar, Germany) and contrasted with 2% uranyl acetate. Detergent treatment was performed before adsorption with 0.1% sodium deoxycholate (Na-DOC) for 10 min at 4 °C as described [25 (link)], to either remove the quasi envelope or to eliminate the virus-like particles of hepatitis B virus surface antigen. For immune-detection, an HEV capsid protein-specific monoclonal antibody [26 (link)] was used together with a gold-labeled secondary antibody. The samples were examined using a JEM 1400 transmission electron microscope (JEOL GmbH, Freising, Germany) operated at 120 kV. Imaging was performed using a Veleta G2 camera (EMSIS GmbH, Münster, Germany). Particles size measurement was done using ITEM software (Olympus, Hamburg, Germany).

A total of 10 µL of cell culture supernatant was adsorbed on to 400 mesh carbon-formvar coated copper grids (Plano GmbH, Wetzlar, Germany) for 5 min followed by inactivation with glutaraldehyde for a further 1 min. Excess liquid was removed by passive capillary action using a tissue paper. The grids were then contrasted with 2% uranyl acetate for 1 min and excess liquid was removed as before. The grids were allowed to dry and examined in a Jeol 1400 Plus TEM (Jeol, Tokyo, Japan), operated at 120 kV. Six different areas of the grid were examined to check the homogeneity of sample. Imaging was performed with Olympus Veleta G2 camera (EMSIS, Muenster, Germany). Particle diameter was measured using ITEM software provided by Olympus.