病原线虫在病组织中可通过挑针直接挑取、漏斗分离或将病组织捣碎分离等方法取得活线虫虫体，为了识别常见线虫，对其形态和结构做一般性观察，可以用活体线虫制作临时玻片，为了进一步仔细观察、认识其分类学特征，需将活体线虫杀死和固定，有时还需染色，制成永久性玻片标本。线虫常规脱水剂Ⅱ由甘油、酒精等组成，需与线虫常规脱水剂 Ⅰ (阿拉丁货号: N1529179) 配合使用，12‑18 小时方可完成线虫样本固定之后从脱水到封藏的整个过程。
自备材料
1、固定液：TAF 固定液或 FAA 固定液等
2、玻皿、培养皿、酒精灯、恒温箱等
3、无水甘油、95% 乙醇、稀碘液、线虫常规脱水剂Ⅰ
操作步骤 (仅供参考)
1、挑取一部分线虫放在载玻片上，滴加一滴蒸馏水，置酒精灯上加热杀死线虫，亦可用稀碘液与线虫悬浮液等比例混合，数秒钟即可杀死线虫。
2、用 TAF 固定液或 FAA 固定液固定线虫，建议采用 TAF 固定，可以长期保存而且线虫形态不易发生变化。
3、固定后的样本放入玻皿中，加入 0.5-1ml 线虫常规脱水剂Ⅰ。
4、将其放在盛有过量 95% 乙醇的密闭容器中，35~40℃恒温箱中脱水处理 12h 以上。
5、再向该玻皿加入 0.5ml 线虫常规脱水剂 Ⅱ，将玻皿放在半封闭培养皿中，置于 40℃恒温箱内，直至脱水剂中的酒精完全蒸发 (约 2-3h)。
6、将脱水处理好的线虫封藏在无水甘油内即可长期保存。
注意事项
1、玻片应洁净，无油污。
2、杀死线虫时不宜用高热火焰。
3、线虫的固定建议采用TAF 固定，可以长期保存而且线虫形态不易发生变化。
4、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Living nematodes can be isolated from diseased tissues by direct picking with a dissecting needle, funnel separation, or homogenization of diseased samples. For general morphological observation and identification of common nematodes, temporary slides can be prepared using live nematodes. To further examine their taxonomic features in detail, living nematodes need to be killed and fixed; staining may be performed as required to prepare permanent glass‑mounted specimens.
Nematode Conventional Dehydrating Solution Ⅱ consists of glycerol, ethanol and other components. It must be used together with Nematode Conventional Dehydrating Solution Ⅰ (Aladdin Cat.No.: N1529179). The whole procedure of dehydration to mounting for nematode samples after fixation takes 12‑18 hours.
Required Materials
1. Fixative: TAF fixative, FAA fixative or equivalent reagents
2. Glass dish, Petri dish, alcohol lamp, incubator, etc.
3. Anhydrous glycerol, 95% ethanol, dilute iodine solution, Nematode Routine Dehydrating Agent Ⅰ
Operating Procedure (For Reference Only)
1. Transfer several nematodes onto a glass slide and add one drop of distilled water. Kill the nematodes by heating over an alcohol‑lamp flame; alternatively, mix the nematode suspension with an equal‑volume of dilute iodine solution for several seconds to kill the specimens.
2. Fix nematodes with TAF fixative or FAA fixative. TAF fixative is recommended for long‑term preservation with minimal morphological distortion.
3. Place the fixed nematode sample into a glass dish and add 0.5‑1 ml of Nematode Routine Dehydrating Agent Ⅰ.
4. Put the glass dish inside a sealed container containing excess 95% ethanol (the sample shall not contact the ethanol liquid). Perform dehydration in an incubator at 35‑40 ℃ for no less than 12 h.
5. Add 0.5 ml of Nematode Routine Dehydrating Agent Ⅱ into the glass dish. Place the glass dish in a semi‑closed Petri dish and incubate at 40 ℃ until ethanol in the dehydrating agent is completely evaporated (approximately 2‑3 h).
6. Mount the dehydrated nematodes in anhydrous glycerol for long‑term storage.
Precautions
1. Ensure glass slides are clean and free of oil contamination.
2. Do not use intense flame when killing nematodes.
3. TAF fixative is recommended for nematode fixation, which enables long‑term preservation and maintains stable nematode morphology.
4. For your personal safety, wear a laboratory coat and disposable gloves during operation.
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