Jem 1200 exii electron microscope

Cortical cells were fixed with 3% glutaraldehyde. Following fixation, dehydration was performed in an ethanol gradient: 30-40-50-60-70-80-90-100% ethanol at 4°C. Then, the cells were embedded in a LR White resin and polymerization was carried out at 50 °C. Ultrathin sections of 70-80 nm were cut from the polymer using an Ultracut-Recheirt-Jung and placed on nickel grids for immunogold assay.
The thin sections were washed twice for 2 min with deionized water and two times with PBS with 0.005% Tween20. Sections were then incubated for 30 min with the blocking solution (50 mM glycine, 0.005% Tween20, 0.01% Triton X-100 and 0.1% BSA in PBS) [53 (link)]. After blocking, sections were incubated with the primary antibody: rabbit anti-LC3 (1:500, MBL PD014, Nagoya, Japan). After rinsing three times in PBS with 0.005% Tween20, the sections were incubated overnight at 4 °C with the secondary antibody: donkey anti-rabbit 25-nm gold conjugate (Electron Microscopy Science Aurion #25708). Samples were washed three times with PBS, 0.005% Tween20 and post-fixed in 2% glutaraldehyde in PBS for 10 min. The sections were then rinsed with distilled water twice for 5 min and contrasted with 2% uranyl acetate, rinsed with water, dried and observed under a JEOL JEM 1200 EXII electron microscope.

Transmission electron microscope was used to observe the fine structure of the materials, which cannot be distinguished by an ordinary microscope. Microscopic images of the pure zeolite material were also taken. JEOL’s Jem 1200 Ex II Electron Microscope (JEOL, Tokyo, Japan), a transmission electron microscope, was used for the study. The samples were bombarded with an electron beam of 80 kV energy.