借助立体定位术和荧光界定GFP裸小鼠脑解剖结构

发布时间：2018-06-14 21:19

本文选题：绿色荧光蛋白 + 裸小鼠脑　；参考：《苏州大学》2012年硕士论文

【摘要】：目的:借助立体定位术和荧光来界定GFP裸小鼠的脑解剖结构。方法:对GFP裸小鼠脑分别进行冠状位、矢状位和水平位的冰冻切片，经荧光成像后行尼氏染色做对比观察，结合Paxinos编著的小鼠脑图谱对荧光图像内的解剖结构进行辨认分析。通过立体定位仪定位10只成年GFP裸小鼠的前囟和人字点所在的脑冠状切面，然后进行冰冻切片，比较小鼠个体间以及与小鼠脑图谱间的解剖结构差异。利用小鼠脑切片模具分别对10个离体鼠脑进行冠状位和矢状位上的切片，再进行冰冻切片，，荧光镜下观察，比较个体间差异，并对离体鼠脑结构进行定位。结果:GFP鼠脑的不同结构间存在着荧光色差，借此可辨认出鼠脑的解剖结构：细胞核、尼氏小体密集区及神经纤维分布区为弱荧光信号，细胞核、尼氏小体密集区周围结构，如嗅球的丛状层、小脑的分子层为强荧光信号。切片经尼氏染色后，荧光强度明显减弱，体视显微镜下观察到的脑结构与荧光镜观察的结果基本一致，参照小鼠脑图谱可对荧光图像上的解剖结构进行辨认。对在体鼠脑的立体定位研究发现GFP小鼠间的前囟、人字点处的冠状切面结构无差异，但前囟处的冠状切面结构与图谱中前囟处的冠状切面结构不同。对离体鼠脑的立体定位研究并未发现个体间的脑切片有差异性，利用小鼠切片模具可进行离体鼠脑的解剖结构定位，准确快捷地获得兴趣位点。结论:借助立体定位术和荧光界定GFP裸小鼠的脑解剖结构可做为一个实验平台应用于目前广为开展的荧光示踪实验的解剖定位研究。
[Abstract]:Aim: to determine the brain anatomy of GFP nude mice by stereotactic localization and fluorescence. Methods: frozen sections in coronal, sagittal and horizontal position of the brain of GFP nude mice were made respectively. The anatomical structure of the fluorescence image was identified and analyzed with the help of the mouse brain atlas compiled by Paxinos. The frontal fontanelle of 10 adult GFP nude mice and the coronal section of the human point were located by stereotactic locator, and then frozen sections were made to compare the anatomical structure difference between the mice and the mouse brain atlas. The coronal and sagittal sections of 10 isolated brains were made by using the mouse brain slice mould, and then frozen sections were observed under fluorescence microscope to compare the differences between individuals, and to locate the brain structure in vitro. Results there were fluorescence color aberrations among the different structures of the brain. The anatomical structure of the brain could be identified: the nucleus, the dense area of Nissl body and the distribution of nerve fiber were weak fluorescence signals, the nucleus, the structure around the dense area of Nissl corpuscles. The molecular layer of the cerebellum, such as the plexiform layer of the olfactory bulb, is highly fluorescent. The fluorescence intensity of the slices was obviously weakened after stained by Nissl, and the observed brain structure under stereoscopic microscope was basically consistent with that observed by fluorescence microscope. The anatomical structure of the fluorescence image could be identified by referring to the mouse brain map. The stereotactic study of the brain in vivo showed that there was no difference in the structure of the coronal section between the GFP mice and the herringbone, but the coronal section structure at the anterior fontanel was different from that at the middle fontanel of the map. The stereotactic localization of the isolated brain has not found any difference among individuals. The mouse slice mould can be used to locate the anatomical structure of the brain in vitro and to obtain the site of interest accurately and quickly. Conclusion: stereotactic localization and fluorescence determination of the brain anatomy of GFP nude mice can be used as an experimental platform to study the anatomical localization of fluorescent tracer experiments.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R322

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