7500 electron microscope

All patients underwent a percutaneous renal biopsy. No symptom perirenal haematoma and macroscopic haematuria have been observed in these patients. Each renal sample contained more than 10 glomeruli. The renal biopsy procedure was as follows: the samples were embedded in paraffin and sectioned at 2 µm, followed by hematoxylin-eosin, Masson, periodic acid-Schiff or periodic acid-silver methenamine (PASM) staining. For immunofluorescence (IF), the samples were sectioned at 3 µm using a cryostat, followed by use of a panel of FITC-conjugated rabbit anti-human antibodies to IgG, IgM, IgA, C3, C1q, and κ and λ light chains (polyclonal, Dako Corporation). These samples were also stained with Congo red. The intensity of the immunofluorescence staining was semiquantitatively scored on a scale of 0 to 2+. Immunophenotyping of the lymphomas was performed on frozen sections using the immunoperoxidase and avidin-biotin techniques. The phenotype of the cellular infiltrate was studied using biotinylated anti-CD4, -CD8, -CD20 and -CD68 antibodies (Dako) and visualized using a peroxidase-streptavidin conjugate. Electron microscopy was performed using a Hitachi 7500 electron microscope after routine sections were prepared from renal tissues, followed by double staining with uranyl acetate and lead citrate. Renal biopsies from all patients were reviewed by two renal pathologists.

HCT-116 cells were harvested by trypsinization, washed twice with PBS, and fixed with ice-cold glutaraldehyde [2.5% glutaraldehyde in PBS (pH 7.4)] for 30min. The cells were washed with PBS, fixed in osmium tetraoxide (OsO₄), and embedded in Spurr's Epon. Representative areas were chosen for ultra-thin sectioning and viewed with a Hitachi 7500 electron microscope.

For transmission electron microscopy (TEM), 20-DAP transgenic T₆ and wild-type kernels were fixed with potassium phosphate buffer (1% glutaraldehyde, 4% paraformaldehyde and 5 mM EGTA, pH 6.8) at 4°C overnight and post-fixed in fixation buffer containing 1% osmium tetroxide at 4°C overnight. Fixed slices were dehydrated in an ethanol gradient up to 100%. The samples were embedded in Spurr and LR White resin for ultrathin sectioning. The thin sections (90 nm) were collected on formvar-coated nickel grids, stained with 2% uranyl acetate and 2.66% lead citrate, and rinsed three times in ddH₂O. The sections were visualized using a Hitachi 7500 electron microscope (Hitachi, Tokyo, Japan) operated at 80 kV.
For scanning electron microscopy (SEM), Mature transgenic T₆ and wild-type kernels were dissected, fixed on a brass disk, and covered with gold/palladium by an ion coater (EIKO IB.3, Japan) The central region of the starchy endosperm was observed by SEM (JEOL, Tokyo, Japan).