Tissue samples were preserved in 2% paraformaldehyde and 0.5% glutaraldehyde in 0.05 M cacodylate buffer (pH 7.4) at 4 °C for 4–18 h. Following fixation, the tissues were treated with reduced osmium tetroxide followed by dehydration in ethyl alcohol. The Spurr’s resin (Electron Microscopy Sciences) was prepared and mixed 1:1 with propylene oxide and incubated overnight on a rotator. A change of Spurr’s resin:propylene oxide at 3:1 was made, and samples were infiltrated overnight. The final embedding was completed in pure Spurr’s resin (hard mixture) for 12–18 h and polymerized at 60 °C. Embedded samples were thin-sectioned (60–80 nm thickness) for electron microscopy using a diamond knife (Diatome) on a Reichert Ultracut E and analyzed using a Philips CM-12 transmission electron microscope at 80 kV accelerating voltage. Mineral crystal size and arrangement were studied by tilting the specimen stage through 0–20° of tilt and/or by darkfield imaging [19 (link)]. Individual crystals could be measured in the darkfield mode and by using the 002-diffraction line from selected-area diffraction.
Ultracut e
Manufactured by Philips
The Ultracut E is a laboratory microtome designed for high-quality sectioning of a wide range of materials. It features a durable, stable construction and precise controls for accurate sample preparation.
Automatically generated - may contain errors
2 protocols using ultracut e
1
Ultrastructural Analysis of Mineral Crystals
2
Ultrastructural Analysis of Mineral Crystals
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Tissue samples were preserved in 2% paraformaldehyde and 0.5% glutaraldehyde in 0.05 M cacodylate buffer (pH 7.4) at 4 °C for 4–18 h. Following fixation, the tissues were treated with reduced osmium tetroxide followed by dehydration in ethyl alcohol. The Spurr’s resin (Electron Microscopy Sciences) was prepared and mixed 1:1 with propylene oxide and incubated overnight on a rotator. A change of Spurr’s resin:propylene oxide at 3:1 was made, and samples were infiltrated overnight. The final embedding was completed in pure Spurr’s resin (hard mixture) for 12–18 h and polymerized at 60 °C. Embedded samples were thin-sectioned (60–80 nm thickness) for electron microscopy using a diamond knife (Diatome) on a Reichert Ultracut E and analyzed using a Philips CM-12 transmission electron microscope at 80 kV accelerating voltage. Mineral crystal size and arrangement were studied by tilting the specimen stage through 0–20° of tilt and/or by darkfield imaging [19 (link)]. Individual crystals could be measured in the darkfield mode and by using the 002-diffraction line from selected-area diffraction.
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