论文部分内容阅读
目的:研究获得背侧齿状回健康颗粒细胞的制片方法。方法:成年小鼠。(1)控制切片角度,尽量保留颗粒细胞顶树突的完整沿水平20~30°切片角度获取海马横向(transverse)脑片;(2)注意区分脑片的正、反面选择海马脑片反面的背侧齿状回上片;(3)合并使用其它保护脑片细胞活性的措施,如采用无钠切片液,切片液中添加抗氧化剂等。结果:本方法所得特定角度的海马横向脑片,其上有大量颗粒细胞表现出轮廓清晰、柔和、胞体圆润的健康状态,且记录出颗粒细胞兴奋性微小突触后电流(mEPSCs)的频率和幅度,均较传统海马横向脑片方法有明显增加。结论:适当的切片角度可保持颗粒细胞远端树突末梢的完整,提高细胞的活性,获得大量健康颗粒细胞。提示在制备脑片时要考虑感兴趣细胞整体的走行,保证切片时细胞整体的完整性。
Objective: To study the preparation of dorsal dentate gyrus granulosa cells. Methods: Adult mice. (1) Control slicing angle, try to keep intact granular cell apical dendrites along the level of 20 ~ 30 ° slice angle to obtain horizontal transverse slices of the hippocampus; (2) pay attention to distinguish the positive and negative brain slices of the negative hippocampal slices Dorsal dentate back on the film; (3) the combined use of other measures to protect the activity of brain cells, such as the use of non-sodium slice solution, slice solution, add antioxidants. Results: The lateral hippocampal slices with specific angle obtained by this method showed a large number of granulosa cells showing a clear, soft and round cell health state, and the frequency of granule cell excitatory postsynaptic currents (mEPSCs) and Amplitude, compared with the traditional method of horizontal slice of the hippocampus has increased significantly. Conclusion: The proper slicing angle can maintain the integrity of the distal dendrites of granulosa cells, increase the activity of cells and obtain a large number of healthy granulosa cells. Tip in the preparation of brain slices to be considered when the overall cell line of interest to ensure the integrity of the overall cell slice.