Jem 1200 transmission electron microscope

HEK293 (p2089) and HEK293 (del-BGLF2) cells were transfected with pCMV-Zta to induce the lytic cycle. On day 3 post-transfection, viral particles in the culture media were harvested and were concentrated by ultracentrifugation. The pellet of viral particles was fixed in 2% paraformaldehyde and 2.5% glutaraldehyde for 30 min at 4°C. The samples were washed and post-fixed in 1% osmium tetroxide for 15 min and then stained with 1% uranyl acetate for 1 h at room temperature. Samples were dehydrated using increasing concentrations of ethanol from 50 to 100% and then embedded in Spurr resin. Embedded samples were sliced into thin sections and stained with uranyl acetate and lead citrate. Images of the samples were obtained using JEOL JEM-1200 transmission electron microscope.

The ultrastructure of the leaf cells was measured according to the methods described by Liu et al. [100 (link)], with minor modification. Leaf samples were cut into small pieces (about 1 cm × 1 cm) and fixed in 2.5% glutaraldehyde solution at 4 °C. The samples were washed five times with 0.1 mol L⁻¹ phosphate buffer and fixed overnight in 1% osmic acid at 4 °C. The materials were dehydrated using a series of acetone solutions (50%, 70% and 90%, v/v), for 15 min at each concentration, and then dehydrated with absolute acetone three times, for 15–20 min each time. The materials were then infiltrated with acetone and resin (EPon812) at proportions of 2:1 (v/v) for 0.5 h at room temperature and at proportions of 1:2 (v/v) for 1.5 h at 37 °C, followed by infiltration with 100% resin for 3 h at 37 °C. After separate polymerization at 37, 45, and 60 °C for 24 h in turn, thin sections were created using an ultramicrotome (Reicher-Jung ULTRACUT, Austria) and the sections were double-stained with uranyl acetate–lead citrate. We examined the samples using a model JEM1200 transmission electron microscope (JEOL, Tokyo, Japan).