An acetylcholinesterase method for in toto staining of peripheral nerves.
An acetylcholinesterase method for in toto staining of peripheral nerves.
复制标题
一种用于外周神经整体染色的乙酰胆碱酯酶方法。
DOI：
10.3109/10520297509117028
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发表时间：
1975
期刊：
影响因子：
--
通讯作者：
J. Drukker
中科院分区：
文献类型：
--
作者：
B. Baljet;J. Drukker
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大鼠胃、小肠、子宫、膀胱、阴道、肠系膜、子宫系膜、关节囊、犬胆囊、胆囊管、胆管、幼儿子宫全部染色。
程序
将组织用含有透明质酸酶的盐水灌注，然后固定在Paraplast的平坦层上，并在冷蔗糖甲醛溶液中固定24小时。胃、膀胱和胆囊也是完全固定的。在冷的0.22 M蔗糖的二甲胂酸钠缓冲液（pH 7.2）中冲洗2天。在由60 mM乙酸盐缓冲液pH 5.0或pH 5.6（仅用于人材料）、2 mM碘化乙酰硫代胆碱、15 mM柠檬酸钠、3 mM硫酸铜、0.5 mM K3 Fe（CN）6、5倍10-4 M iso-OMPA、1% Triton X 100组成的培养基中于37 ℃孵育。用双蒸水冲洗。在甘油含量增加的甘油/水混合物中脱水。在甘油中储存或在解剖显微镜下分层。将分层的样本固定在凝胶化的物镜上，在二甲苯中清洁并用Malinol盖上。储存在甘油中的样本可在显微镜下进行研究。染色的标本也可以包埋在Paraplast中，复染后可以研究切片。
UNLABELLED
Stomach, small intestine, uterus, urinary bladder, vagina, mesentery, mesometrium and joint capsule of rats, gall bladder, cystic duct and bile duct of dogs and uteri of young children are stained in toto.
PROCEDURE
Tissue is perfused with saline containing hyaluronidase, then pinned on a flat layer of Paraplast and fixed for 24 hr in cold sucrose formol solution. Stomach, urinary bladder and gall bladder are also fixed in toto. Rinse for 2 days in cold 0.22 M sucrose in a sodium cacodylate buffer pH 7.2. Incubate in medium consisting of 60 mM acetate-buffer pH 5.0 or pH 5.6 (for human material only), 2 mM acetylthiocholine iodide, 15 mM Na citrate, 3 mM Cu sulphate, 0.5 mM K3Fe(CN)6, 5 times 10-4 M iso-OMPA, 1% Triton X 100 at 37C. Rinse in doubly distilled water. Dehydrate in glycerine/water mixtures of increasing glycerine content. Store in glycerine or delaminate under dissecting microscope. Delaminated specimens are mounted on gelatinized object glasses, cleared in xylene and coverslipped with Malinol. Specimens stored in glycerine can be studied microscopically. Stained specimens can also be embedded in Paraplast and sections can be studied after counterstaining.