The amount of time in which to learn the material is fixed, but the amount of learning varies significantly.
The material was fixed at 2.5% glutaldehyde and 2% paraformaldehyde solution; post-fixed at 2% osmium tetroxide; dehydrated with increasing ethanol concentrations, and included in epoxy resin.
For this, the material was fixed in glutaraldehyde 2.5%, soon after it was submerged in buffer phosphate during 24 hours, then fixed in osmium tetroxide (OsO4) 1.5% at 2°C for 3 hours, dehydrated in an upward series of acetone and embedded in Spurr's resin.
The material was fixed in a 10% formalin solution and embedded in paraffin.
In the control sections, where the material was fixed without the addition of pyroantimonate, electron-dense precipitates did not occur.
All the material was fixed and stored in 4 % solution of paraformaldehyde in phosphate-buffered saline (PBS).
After the fixation the material was fixed again for 1 h in 1% solution of osmium quatroxide in the same buffer at 4 °C.
To enable an ultrastructural analysis, the material was fixed in 4% buffered paraformaldehyde, postfixed in 1% OsO4, dehydrated with alcohol in increasing concentrations, and embedded in Epon-Araldite.
Subsequently, the material was fixed in 3 1 v/v absolute ethanol:glacial acetic acid and stored at 4 °C for 24 h, and then stored in 70 % ethanol at 4 °C until use.
After washing in 0.1 M phosphate buffer (pH 7.4), the material was fixed in 1% OsO4/0.1 M phosphate buffer (pH 7.3) at 3 °C for 1½ hours and washed again in 0.1 M phosphate buffer (pH 7.4).
The material was fixed in 2.5 % (w/v) glutaraldehyde in 50 mM cacodylate buffer (pH 7.4) for 2 h, washed in the buffer and placed in a 2%% (w/v) osmium tetroxide at 4°C in 50 mM cacodylate buffer (pH 7.4) for about 12 h.
