Transmission
electron microscopy (TEM) analysis was performed as previously described.19 (link) Briefly, cells were fixed with 3% paraformaldehyde
and 2% glutaraldehyde in 0.1 M cacodylate buffer containing 5 mM CaCl2 (pH 7.4) and then postfixed in 1% osmium tetroxide supplemented
with 0.5% potassium hexacyanoferrate trihydrate and potassium dichromate
in 0.1 M cacodylate for 1 h. The cells were then stained with 2% uranyl
acetate in water for 1 h, dehydrated in graded ethanol solutions,
and embedded in Agar 100 epoxy resin (Agar Scientific Ltd., Stansted,
UK). Ultrathin sections (70–90 nm) were viewed and photographed
with an FEI Tecnai SPIRIT (FEI, Eidhoven, Netherlands) transmission
electron microscope operated at 120 kV and equipped with a OneView
Gatan camera.
electron microscopy (TEM) analysis was performed as previously described.19 (link) Briefly, cells were fixed with 3% paraformaldehyde
and 2% glutaraldehyde in 0.1 M cacodylate buffer containing 5 mM CaCl2 (pH 7.4) and then postfixed in 1% osmium tetroxide supplemented
with 0.5% potassium hexacyanoferrate trihydrate and potassium dichromate
in 0.1 M cacodylate for 1 h. The cells were then stained with 2% uranyl
acetate in water for 1 h, dehydrated in graded ethanol solutions,
and embedded in Agar 100 epoxy resin (Agar Scientific Ltd., Stansted,
UK). Ultrathin sections (70–90 nm) were viewed and photographed
with an FEI Tecnai SPIRIT (FEI, Eidhoven, Netherlands) transmission
electron microscope operated at 120 kV and equipped with a OneView
Gatan camera.
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