Electron microscopy was performed as described previously [47 ]. NRVMs were seeded on gelatin-covered 35 × 10 mm Permanox dishes (Electron Microscopy Sciences, 70,340) at a density of 2 × 105 cells per plate and transfected with siRNA as described above. Six days after transfection, NRVMs were rinsed with warm (room temperature), diluted 1:1 with ddH2O, cardioplegic buffer (50 mmol/L KCl, 5% dextrose in PBS, pH 7.4) and fixed on ice in 1% paraformaldehyde/2% glutaraldehyde in 50 mM cacodylate buffer, pH 7.2, for 20 min, postfixed in reduced osmium (1% OsO4, 0.75% K3Fe(CN)6, 25 mM cacodylate buffer, pH 7.2), dehydrated using a graded series of acetone, and embedded in epoxy resin. After polymerization thin layer containing cell monolayers were detached from the bottom of the dish, cut into small fragments and glued to stubs for sectioning. Ultrathin sections were counterstained with uranyl acetate and lead salts. Images were acquired on a Hitachi 7600 electron microscope equipped with an Advanced Microscopy Techniques digital camera.
7600 electron microscope
Manufactured by Hitachi
Sourced in Japan
The Hitachi 7600 electron microscope is a high-performance imaging system designed for advanced materials analysis. It features a stable electron beam and precision optics to provide clear, high-resolution images of samples. The 7600 is capable of magnifying specimens up to 1,000,000x, enabling detailed examination of microstructures and nanoscale features.
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Lab products found in correlation
5 protocols using 7600 electron microscope
1
Electron Microscopy of Cardiomyocytes
2
Visualizing CaP-rHDL Uptake in Glioblastoma
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To examine the ultrastructure of cellular uptake of CaP-rHDL in C6 glioblastoma
cells, the cells were incubated with CaP-rHDL or CaP-LNC for 3 h at the
DMPC concentration of 100 μg ml−1. After
that, the cells were washed with PBS for two times, fixed with 2.5%
glutaraldehyde at 4 °C for 2 h, scratched off from the flask,
centrifuged at 1,500 r.p.m. for 5 min with the pellet re-suspended
in 2.5% glutaraldehyde, stored at 4 °C until post-fixation in
1% OsO4 in 1 M PB, and finally subjected to ultra-thin
section and microscopic analysis under a Hitachi 7600 electron microscope.
cells, the cells were incubated with CaP-rHDL or CaP-LNC for 3 h at the
DMPC concentration of 100 μg ml−1. After
that, the cells were washed with PBS for two times, fixed with 2.5%
glutaraldehyde at 4 °C for 2 h, scratched off from the flask,
centrifuged at 1,500 r.p.m. for 5 min with the pellet re-suspended
in 2.5% glutaraldehyde, stored at 4 °C until post-fixation in
1% OsO4 in 1 M PB, and finally subjected to ultra-thin
section and microscopic analysis under a Hitachi 7600 electron microscope.
3
Electron Microscopic Analysis of AZD2281 and ATG5 Knockdown
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Cells were grown on 6-well plates, treated with AZD2281, ATG5 shRNA, or control shRNA, fixed for 2 h with 2.5% glutaraldehyde in 0.1 mol/l cacodylate buffer (pH 7.4), and postfixed in 1% OsO4 in the same buffer and then subjected to the electron microscopic analysis as described previously. Representative areas were chosen for ultrathin sectioning and viewed with a Hitachi 7600 electron microscope (Japan).
4
Electron Microscopy Analysis of MOECs
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MOECs were fixed for 2 hours with 2.5% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.4), fixed again in 1% OsO4 in the same buffer, and then subjected to electron microscopy analysis as described previously (Klionsky et al., 2008 (link)). Ultra-thin sections in representative areas from five fields are shown. Sections were viewed with a Hitachi 7600 electron microscope (Hitachi High Technologies America, Inc., Pleasanton, CA). The substructure was observed under 5000× and 50,000× magnification.
5
Electron Microscopy Analysis of MOECs
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