The germinated seeds were first fixed in a 2.5% glutaraldehyde fixation solution in 0.1 M Na-phosphate butter for 2 h at room temperature and then held at 4 °C. The seeds were washed 3 times with phosphate buffer, successively dehydrated in gradient ethanol, and embedded in LR White Resin. The semithin sections of 2 μm thickness were cut with a glass knife on a Leica Ultrathin Microtome (EM UC7) following the method of Zhao et al. [21 (link)]. The sections were counterstained with periodic acid-Schiff reagent and toluidine blue O, and viewed with an Olympus BX53 light microscope equipped with a CCD camera.
Ultrathin microtome em uc7
Manufactured by Leica
Sourced in Germany
The Leica Ultrathin Microtome (EM UC7) is a precision instrument designed for the sectioning of biological samples. It allows for the cutting of extremely thin sections, typically in the range of 50 to 500 nanometers, for examination under an electron microscope. The EM UC7 features a high-stability cryogenic system, ensuring accurate and consistent sectioning at low temperatures.
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Lab products found in correlation
2 protocols using ultrathin microtome em uc7
1
Seed Microstructure Analysis Protocol
2
Grain Sample Preparation for Microscopy
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Grains samples were fixed immediately in 0.1 M phosphate buffer solution (pH 7.2), containing 2.5% glutaraldehyde (v/v) and 2% paraformaldehyde (w/v). Afterwards, they were dehydrated in a graded ethanol series, transferred to acetone, and embedded in low-glutinosity Spurr's resin (Spurr Low-Viscosity Embedding Kit, Sigma-Aldrich, St. Louis, USA). The sample blocks were polymerized at 70°C for 8 h. Semi-thin sections (2 μm) were cut at the longitudinal plane (embryo) and the transversal plane (endosperm) of the grains, using a Leica Ultrathin Microtome (EM UC7, Germany). Samples were stained with 1 % methyl violet, observed and photographed under a BX53 light microscope (Olympus, Tokyo, Japan).
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