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背景:在神经损伤与修复的研究工作中,常常会涉及损伤后以及修复过程中神经元和神经纤维的定性、定位和半定量的问题。目前神经生物学常规实验技术还很难解决这样的问题。目的:探索一种简便易行并能同时显示大鼠中枢神经系统(centralner-voussystem,CNS)神经元和神经纤维的全程连续切片的块染方法。设计:非随机非对照的实验研究。地点和材料:第三军医大学组胚教研室(组织学常规技术实验室),实验动物中心。动物:健康成年Wistar无菌级大鼠2只(第三军医大学实验动物中心提供),体质量200～300g,雌雄不拘。干预:取大鼠嗅脑至脊髓骶段的完整CNS组织,将CNS按顺序以冠状切面切成厚2cm的块待切,氨乙醇固定组织吸干后直接入30g/L硝酸银水溶液,于22℃恒温箱中浸银染色1周。常规脱水、透明、石蜡包埋,全程连续切片的制作。一个大鼠的CNS全程标本可做成3套相近的连续切片。主要观察指标:神经元及突起和神经纤维的形态结构。结果:大鼠全程CNS连续切片各断面上神经元呈深棕黄色,核膜及核仁呈黑色;神经纤维呈黑色;背景淡黄色。结论:运用灌流冲洗后固定、Cajal氏镀银块染、连续石蜡切片的方法,可制作同时显示CNS神经元和神经纤维的全程连续切片。将此技术方法运用到神经损伤与修复的实验研究中,对于神经元和神经纤维的定性
BACKGROUND: In the research of nerve injury and repair, the problems of qualitative, localization and semi-quantification of neurons and nerve fibers after injury and repair are often involved. At present, conventional neurobiology experiments are still very difficult to solve such problems. OBJECTIVE: To explore a simple and easy method for the simultaneous lumping of serial sections of rat central nervous system (CNS) neurons and nerve fibers. Design: A Non-randomized, Uncontrolled Experimental Study. Location and Materials: Department of Embryology, Third Military Medical University (Histology and Conventional Technology Laboratory), Laboratory Animal Center. Animals: 2 healthy adults Wistar sterile rats (Experimental Animal Center, Third Military Medical University), body weight 200 ~ 300g, male or female. INTERVENTIONS: The intact CNS tissues of the rat sniffing brain to the sacral segment of the spinal cord were taken. The CNS was sequentially cut in a coronal section into 2 cm thick blocks to be cut. The fixed ammonia-ammonia solution was directly drained into 30 g / L silver nitrate aqueous solution and treated at 22 Celsius incubator immersion silver staining for 1 week. Conventional dehydration, transparent, embedded in paraffin, the entire production of serial sections. A rat CNS full specimen can be made into three sets of similar serial sections. MAIN OUTCOME MEASURES: Morphological structure of neurons and protrusions and nerve fibers. RESULTS: The neurons in each section of CNS serial sections of rats were dark brown, nuclear membrane and nucleolus were black, nerve fibers were black, and the background was yellowish. CONCLUSIONS: Serial perfusion of CNS neurons and nerve fibers can be made simultaneously by perfusion-flushing, fixation with Cajal silver stain, continuous paraffin sectioning. Applying this technical method to the experimental study of nerve injury and repair, the qualitative analysis of neurons and nerve fibers