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目的:通过冰冻切片和脑片培养方式比较获得更适合脑片实验研究的方法。方法:分别运用急性切片和脑片培养方法,结合全细胞膜片钳技术比较两种脑片处理方法对小鼠海马神经元细胞形态、细胞膜封接难易程度、细胞电生理特性等的差异,获得更适合细胞研究的脑片获取方法。结果:冰冻切片方法切断部分神经纤维,脑片表层出现肿胀或坏死细胞,2-3层细胞可用于膜片钳记录,但不易封接破膜。脑片培养后可使纤维再生,整个脑片细胞形态清晰可见,容易封接破膜,电生理记录波形及基本特性与冰冻切片一致,但脑片培养方法的细胞突触后电流幅度更大、频率更高。结论:脑片培养可修复受损纤维和细胞膜柔韧性,且不改变膜特性,但脑片培养重建了一定数量的细胞间信号连接,使细胞反应性增强,脑片培养方法更适合脑片神经元研究。
OBJECTIVE: To compare the methods of frozen slice and slice culture to get more suitable brain slices. Methods: Acute and sliced slices were used in combination with whole-cell patch-clamp technique to compare the differences of neuronal cell morphology, ease of cell membrane sealing and electrophysiological characteristics in hippocampus of mice treated by two kinds of brain slices. More suitable for cell research methods to obtain brain slices. Results: Frozen section method cut off part of the nerve fibers, the brain surface swelling or necrosis of cells, 2-3 layers of cells can be used for patch-clamp recording, but not easy to seal rupture of membranes. After cultured, the regenerated fibers could be regenerated. The morphology of the whole brain slices was clearly visible. It was easy to seal the rupture membrane. The electrophysiological recording waveform and the basic characteristics were consistent with frozen sections. However, Higher frequency. CONCLUSION: Brain slice culture can repair the damage of damaged fiber and cell membrane, and does not change the membrane properties. However, the number of cells in brain slice reconstructs a certain number of signal connections and enhances cell reactivity. The method of brain slice culture is more suitable for cranial nerves Meta research.