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目的筛选适宜吸虫神经系统的染色方法。方法取河蚌体内自然感染的贝居腹盾吸虫并分为2组,先用10%甲醛固定,再分别用乙酰胆碱酯酶组织化学定位方法和Cajal镀银染法对吸虫进行整体染色,经逐级乙醇脱水、透明、中性树胶封片。显微镜下观察神经中枢、神经纤维、神经细胞,对比2种染色方法的优缺点,同时对染液配制、染色步骤、染色条件进行比较。结果乙酰胆碱酯酶组化染色法能清晰显示贝居腹盾吸虫神经整体结构,包括中枢神经节、主要神经干、环腹吸盘的神经环、体表神经纤维等,而生殖器官、消化器官神经分布显示不充分;Cajal银染法可显示出吸虫体表及内脏器官分布的神经纤维,局部可以显示神经细胞,但不能显示较为粗大的神经干,此法染色对比度较差。乙酰胆碱组织化学方法染液配制较复杂,染液有效期短,仅约3h,但染色步骤简单;银染法染液可以重复使用,但染色步骤复杂,可能受多种因素影响。结论观察吸虫清晰的神经整体结构可用乙酰胆碱组织化学法,观察神经纤维及神经元结构则可以采用银染法;两种染色方法各有侧重,结合使用有助于全面了解吸虫神经系统解剖结构。
Objective To screen the suitable method for staining the nervous system of fluke. Methods The mussels were exposed to the natural infection of mussels. The animals were divided into 2 groups, fixed with 10% formalin, and then stained with acetylcholinesterase histochemical method and Cajal silver staining, respectively. Ethanol dehydration, transparent, neutral gum seal. The nerve center, nerve fibers and nerve cells were observed under the microscope. The advantages and disadvantages of the two staining methods were compared. Meanwhile, the preparation of the dye solution, the dyeing procedure and the dyeing conditions were compared. Results Acetylcholinesterase histochemical staining could clearly show the whole structure of the neurotrophic nerve, including the central nervous system, the main nerve trunk, the ring of the supine abdominal suction, and the surface nerve fibers, while the reproductive organs and digestive organs Showing inadequate; Cajal silver staining method can be displayed on the body surface and visceral organs of parasitic nerve fibers, nerve cells can be displayed locally, but can not show a thicker nerve trunk, this method staining contrast is poor. Acetylcholine histochemical staining method is more complex preparation, the dye is short, only about 3h, but the dyeing step is simple; silver staining dye can be reused, but the dyeing step is complicated and may be affected by many factors. Conclusion The clear structure of nerve can be treated with acetylcholine histochemistry, and silver staining can be used to observe the structure of neurofibers and neurons. The two staining methods have their own focus, which helps to understand the anatomical structure of the nervous system.