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目的:通过改变传统的Golgi-Cox染色方法,探究一种在海马区神经元形态与结构的研究中更加稳定和有效的Golgi-Cox染色法。方法:本实验将昆明小鼠随机分为两组,一组采用传统的Golgi-Cox染色法,另一组采用本研究中改良的Golgi-Cox染色法。改良方法在传统方法的基础上,改变了以下几个关键环节,包括固定、切片、脱水和显色等,然后对两种方法的结果进行差异比较;为减少资料分析上的偏差,由资料分析人员对实验结果进行统计学分析。结果:改良法比较传统法,通过在离心管内悬空固定组织,切片时用琼脂包埋,延长低浓度乙醇脱水及二甲苯透明时间,并缩短高浓度乙醇脱水时间,改善显色环境的方法,可以在更好地保持组织切片的完整性,减少脱片的情况下,有效减少背景染色,增强显色效果。稳定地显示出更多的胞体(增加50%)、树突分支(增加79%)和树突棘(增加42%)。结论:改良的Golgi-Cox染色方法比传统的Golgi-Cox染色方法更加稳定和敏感,在海马区神经元树突和树突棘形态与结构研究中是一种可靠的技术方法。
OBJECTIVE: To explore a more stable and effective Golgi-Cox staining method in the study of neuronal morphology and structure in hippocampus by changing the traditional Golgi-Cox staining method. Methods: Kunming mice were randomly divided into two groups, one group using the traditional Golgi-Cox staining method and the other group using the modified Golgi-Cox staining method in this study. Based on the traditional method, the improved method changes the following key aspects, including fixing, slicing, dehydration and color development, and then compares the results of the two methods; in order to reduce the deviation of data analysis, The staff of the experimental results for statistical analysis. Results: Compared with the traditional method, the modified method was able to immobilize the tissue in the centrifuge tube, embedding with agar when slicing, prolonging the time of low concentration ethanol dehydration and xylene clearing, shortening the time of high concentration ethanol dehydration, and improving the color developing environment. In the better to maintain the integrity of tissue sections, reduce the case of off-chip, effectively reduce the background staining, enhance the color effect. Stable display of more somatic cells (50% increase), dendritic branches (79% increase) and dendritic spines (42% increase). CONCLUSION: The modified Golgi-Cox staining method is more stable and sensitive than the traditional Golgi-Cox staining method. It is a reliable technique in the study of morphology and structure of dendrites and dendritic spines in the hippocampus.