Live nematode specimens can be obtained from infected tissues via direct picking with a needle, Baermann funnel extraction, or grinding the infected tissues followed by separation. For the identification of common nematode species, general observations of their morphology and structure can be conducted by preparing temporary slides using live nematodes. To further examine and characterize their taxonomic features in detail, live nematodes need to be killed, fixed, and sometimes stained to prepare permanent slide specimens. If the goal is to observe the interaction between nematodes and host tissues, paraffin sections of both host and nematode tissues are required.
Aladdin Nematode Vital Stain Solution (Iodine Method) is mainly composed of iodine, potassium iodide, and other ingredients. It is designed for the vital staining and observation of plant tissue samples containing nematodes, and is particularly suitable for root-inhabiting nematode materials. Its working principle relies on the penetration of iodine solution into root tissues, which stains the nematodes while leaving the plant tissues unstained, thus facilitating the distinction between the two. This reagent is for research use only and is not intended for clinical diagnosis or any other purposes.
Self-prepared Materials:
Distilled water, glass slides, microscopes, tweezers, dissecting needles, glycerol, mounting media, etc.
Operating Procedures (For Reference Only):
1. Rinse the diseased roots of plants with distilled water and blot the water dry with absorbent paper.
2. Immerse the diseased roots in iodine solution with tweezers and stain for 10–15 minutes.
3. Take out the tissues with tweezers and rinse them with distilled water.
4. Place the tissues on a glass slide, locate the nematodes with a dissecting needle, and observe the staining status of the tissues and nematodes.
5. If necessary, squash the preparation to observe the staining of the internal organs of the nematodes.
6. Prepare temporary or permanent mounts using Arfagal mounting medium.
Staining Results:
Nematodes
| Brownish yellow to black
|
Pathological samples
| Colorless
|
Preautions:
1. Glass slides shall be clean and free of oil stains.
2. Stained nematode specimens can be preserved in anhydrous glycerol for 2–5 months, after which they will gradually fade.
3. Under normal circumstances, nematodes will be stained brownish yellow to black, but not all morphological characteristics can be visualized by staining. That is to say, the staining results may not be completely consistent with the table above.
4. High-temperature flame fixation should be avoided during the fixation process.
5. For your safety and health, please wear a lab coat and disposable gloves when performing the operation.
6. Please use the reagent as soon as possible after opening to prevent affecting the subsequent experimental results.
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