菌根侵染状况的观察和侵染率的测定，可以用活体镜检法和染色镜检法。活体镜检法的优点是可连续活体观察多次，能及时大体了解菌根发育情况，并可节省植株。缺点是并不十分准确。为了精确检查菌根发育状况、测定侵染率，则需采用染色法。一般要经过根系透明-染色-分色过程。
阿拉丁乳酸台盼蓝染色脱色液主要由乳酸、甘油、水等组成，用于经乳酸台盼蓝染色后的脱色。本产品可用于霉菌的染色观察，也可用于丛枝菌根的染色研究。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
自备材料：
1、蒸馏水、FAA固定液、5%~10%KOH溶液、2%的HCl溶液、乳酸台盼蓝染色液(0.05%)
2、显微镜、、水浴锅
操作步骤(仅供参考)：
1、将根段或经过FAA固定的根系剪成长0.5~1.0cm的小段放入试管或其他染色容器，加入5%~10%KOH溶液，放在90°C水浴锅内10~20min。
2、弃液，流水冲洗。加入碱性过氧化氢，室温下放置15min后弃液，流水冲洗。
3、加入2%的HCl溶液，浸没根系，室温下放置5~20min后弃液，流水冲洗。
4、加入乳酸台盼蓝染色液(0.05%)，置于90°C水浴锅内染色30~60min或室温过夜染色。
5、弃液，流水冲洗。加入乳酸台盼蓝染色脱色液，室温下脱色1~2天，除菌根中AMF有蓝色以外，根系中无多余的颜色。
6、玻片上滴加甘油，放上根段后封片镜检。
染色结果：
植物细胞不着色(有时中柱着色)，真菌组织染成蓝色。
注意事项：
1、通常草本植物的幼嫩根系透明时间较短，而木本植物的根系则较长。
2、乳酸台盼蓝染色液(0.05%)可回收重复再用。
3、为了您的安全和健康，请穿实验服并戴一次性手套操作。
4、试剂开封后请尽快使用，以防影响后续实验效果。
The observation of mycorrhizal colonization and the determination of colonization rate can be carried out by the vital microscopic examination method and the staining microscopic examination method. The advantage of the vital microscopic examination method is that it allows repeated continuous observation of living samples, enables timely and general understanding of mycorrhizal development status, and helps save plant materials. Its disadvantage is that the results are not very accurate. To accurately examine mycorrhizal development and determine the colonization rate, the staining method should be adopted, which generally involves the processes of root clearing, staining and differentiation.
Aladdin Lactate Trypan Blue Staining and Destaining Solution is mainly composed of lactic acid, glycerol and water. It is used for destaining after staining with lactate trypan blue staining solution. This product can be applied to the staining and observation of molds, and also to the staining research of arbuscular mycorrhizae. This reagent is for research use only and is not intended for clinical diagnosis or any other purposes.
Self-prepared Materials:
1. Distilled water, FAA fixative, 5%–10% KOH solution, 2% HCl solution, 0.05% lactate trypan blue staining solution
2. Microscope, water bath
Operating Procedures (For Reference Only):
1. Cut root segments or roots fixed with FAA into small pieces of 0.5–1.0 cm in length, place them into a test tube or other staining container, add 5%–10% KOH solution, and incubate in a water bath at 90 °C for 10–20 minutes.
2. Discard the solution and rinse thoroughly under running water. Add alkaline hydrogen peroxide, leave it at room temperature for 15 minutes, then discard the solution and rinse again under running water.
3. Add 2% HCl solution to submerge the roots, incubate at room temperature for 5–20 minutes, discard the solution and rinse under running water.
4. Add 0.05% lactate trypan blue staining solution, and stain either in a water bath at 90 °C for 30–60 minutes or overnight at room temperature.
5. Discard the staining solution and rinse under running water. Add lactate trypan blue staining and destaining solution, and destain at room temperature for 1–2 days until only the AMF in the mycorrhizae shows blue coloration, with no excess color remaining in the root tissues.
6. Place a drop of glycerol on a glass slide, mount the root segments, seal the slide and observe under a microscope.
Staining Results:
Plant cells remain unstained (sometimes the stele is stained), while fungal tissues are stained blue.
Precautions:
1. Generally, the clearing time required for the tender roots of herbaceous plants is relatively short, whereas that for the roots of woody plants is longer.
2. The 0.05% lactate trypan blue staining solution can be recovered and reused.
3. For your safety and health, please wear a lab coat and disposable gloves during the operation.
4. Please use the reagent as soon as possible after opening to avoid affecting the results of subsequent experiments.
ZJ26F0232110
危险品化学品经营许可证（带存储）