H7700 electron microscope

For transmission electron microscopy (TEM), viral particles from infected alfalfa tissues were partially purified using a protocol developed for Poinsettia mosaic virus [15 ]. For TEM observation of virus-like particles generated via the PVX vector in N. benthamiana plants, samples were processed as described in [6 (link)]. Virus captured on the TEM grids was stained with 1% phosphotungstate (PTA) solution. The grids were examined in a Hitachi H-7700 Electron Microscope at the Electron and Confocal Microscope Unit, Beltsville Agricultural Research Center.

For TEM, NPCs infected with S. aureus on chamber slides and the caudal rat IVDs infected with S. aureus were fixed with 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.4) for 2 h, respectively. Conventional electron microscopy was performed as follows. After five washes with 0.1 M phosphate buffer, the cells were post-fixed with 2% osmium tetroxide and 0.5% potassium ferrocyanide in the same buffer for 1 h and then washed again with 0.1M phosphate buffer. After dehydration, the cells and tissues were embedded in Epon 812 (TAAB Laboratories Equipment Ltd.). Ultrathin sections were stained with uranyl acetate plus lead citrate and observed using an H7700 electron microscope (Hitachi, Tokyo, Japan).

HK-2 cells were treated with MA and vehicle separately. The cells were digested and collected routinely. The cells were suspended and fixed with 2.5% glutaraldehyde in 0.1 mM PBS (pH 7.4) at 4 °C for 2 h. After washing twice with PBS, the cells were treated with conventional dehydration, osmosis, embedding, sectioning, and staining. The ultrastructure of the cells was observed under a Hitachi H7700 electron microscope.

Mouse hippocampal tissue was preserved using an electron microscopy fixative (Servicebio, G1102) before undergoing a series of standard histopathological steps, including dehydration, osmication, embedding, sectioning, and staining. The ultrastructural details of the hippocampal neurons were then closely examined using a Hitachi H7700 electron microscope.

The fixed procedure was as described^{16 (link)}. Samples were immersed with 2% paraformaldehyde and was further fixed with sequential incubation with 1 and 2% glutaraldehyde in PBS at 4 °C for 24 h. Post-fixed procedure was provided with 1% osmium tetroxide, electron-stained with 3% uranyl acetate, and embedded in an Epon-Araldite mixture. Then ultrathin sections were cut with approximately 60 nm by a ultrathin microtome (Leica EM UC7) and detected by a Hitachi H-7700 electron microscope (Hitachi, Tokyo, Japan).

For electron microscopy, cells infected with P. acnes on chamber slides were fixed with 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.4) for 2 h. Conventional electron microscopy was performed as follows. After washing five times with 0.1 M phosphate buffer, the cells were post-fixed with 2% osmium tetroxide and 0.5% potassium ferrocyanide in the same buffer for 1 h, and washed again with 0.1 M phosphate buffer. After dehydration, the cells were embedded in Epon 812 (TAAB Laboratories Equipment Ltd.). Ultrathin sections were stained by uranyl acetate plus lead citrate and observed with an H7700 electron microscope (Hitachi, Tokyo, Japan).

HK-2 cells were treated as mentioned above. Then, the cells were collected and undergone dehydration, osmosis, embedding, sectioning, and staining, as previously described [7 (link)]. Typical images were captured by a Hitachi H7700 electron microscope.