BioReagent BioReagent for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
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Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
Semi-thin sectioning is a plastic sectioning technique used to prepare specimens for observation and study under a light microscope. The procedure is the same as that for ultrathin sectioning (section thickness below 0.1 μm, usually 60–80 nm), but semi-thin sections are approximately 0.5–1.5 μm thick, intermediate between paraffin sections and ultrathin sections. The results are better than those of paraffin sections because the sections are thinner, allowing better demonstration of cellular structures and reducing artifacts; the imaging quality is clearer than that of paraffin sections. It is often used for the observation of nerve tissue sections.
Semithin section staining solution (Toluidine blue method) consists of toluidine blue, borate, deionized water, etc. Because borate buffer is strongly alkaline, it is more conducive to staining tissue cells. Transverse semi-thin sections stained with toluidine blue can be used to observe lesions of the nerve myelin sheath and myelin thickness under a light microscope; they can also be used for plant leaves, anthers, etc., to observe cell arrangement, etc. This reagent is for research use only and is not suitable for clinical diagnosis or other purposes.
Operating Procedures (For reference only):
1. Distilled water or deionized water, graded ethanol, 1% osmic acid fixative, 0.1 M PB phosphate buffer (pH 7.4)
2. Acetone, 812 embedding medium, xylene or environmentally friendly dewaxing clearing solution, neutral balsam
3. Embedding plates, oven, semi-thin microtome, glass slides, microscope
Procedure (for reference only):
1. Fix tissue in 1% osmic acid prepared in 0.1 M PB phosphate buffer (pH 7.4) at room temperature in the dark for 7 h. Rinse with 0.1 M PB phosphate buffer (pH 7.4) 3 times, 15 min each.
2. Dehydrate at room temperature: sequentially transfer tissue into 30%, 50%, 70%, 80%, 95%, 100%, and 100% ethanol for dehydration, 1 h each.
3. Absolute ethanol:acetone (3:1) for 30 min, absolute ethanol:acetone (1:1) for 30 min, absolute ethanol:acetone (1:3) for 30 min, acetone for 1 h.
4. Infiltration: acetone:812 embedding medium (3:1) at 37°C for 2–4 h; acetone:812 embedding medium (1:1) at 37°C overnight; acetone:812 embedding medium (1:3) at 37°C for 2–4 h; pure 812 embedding medium at 37°C for 5–8 h.
5. Embedding: Pour pure 812 embedding medium into embedding plates, insert samples into the embedding plates, and embed overnight in a 37°C oven.
6. Polymerization: Place the embedding plates in a 60°C oven for polymerization for 48 h, then remove the resin blocks for later use.
7. Sectioning: Section with a semi-thin microtome at a thickness of 1.5 μm, and attach the tissue sections to glass slides.
8. Staining: Heat the toluidine blue staining solution in a 60°C oven for 1 h, then place the slides in the staining solution for 2 min.
9. Wash with water, differentiate in 95% ethanol, control the color under a light microscope, dry in an oven, mount with neutral balsam, and perform whole-slide scanning.
Precautions：
1. The embedding surface of the specimen should not be too large; otherwise, wrinkles are easily formed.
2. Staining should preferably be carried out under heating to facilitate penetration of the dye into the resin.
3. Semi-thin sections can be examined by phase contrast, or observed under a light microscope after staining.
4. Glass knives should be used for semi-thin sectioning; do not use diamond knives.
5. Osmium tetroxide (osmic acid) vapor also has a tissue-fixing effect. Therefore, when preparing osmic acid fixative, wear gloves and goggles for protection. Specimens fixed with aldehyde must be thoroughly washed with buffer before entering osmic acid to prevent interaction between the two fixatives.
6. For your safety and health, please wear a lab coat and disposable gloves during operation.
Comprehensive hazard, handling, storage, and regulatory compliance document.
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