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Abstract：
The localization areas of intracellular proteins in rat cortical neurons were visualized using a scanning electron microscope (SEM) coupled with a focused ion beam (FIB) system. To obtain a clear contrast in the SEM images, gold nanoparticles (GNPs) were bound to specific intracellular proteins by antigen-antibody reactions. By obtaining a cross section of the desired location of the neurons by FIB milling under the SEM imaging condition, it was possible to observe the proteins inside the cells as clear bright spots. When a neuron was stained with anti-tau and anti-histone H1 antibodies, the bright spots were localized in the cross section of the axon and the nucleus, respectively. It was confirmed that targeted proteins in a single neuron on a substrate could be successfully identified. The development of FIB/SEM observation with immunological GNP staining will offer important information for the stable growth of neurons on various substrate structures, since the elongation and turning of axons on the substrates are activated by the redistribution of intracellular proteins.
摘要：
使用扫描电子显微镜（SEM）和聚焦离子束（FIB）系统可视化大鼠皮质神经元中细胞内蛋白质的定位区域。为了在SEM图像中获得清晰的对比度，金纳米颗粒（GNP）通过抗原-抗体反应与特定的细胞内蛋白结合。通过在SEM成像条件下通过FIB铣削获得神经元所需位置的横截面，可以观察到细胞内的蛋白质是清晰的亮点。当神经元用抗tau和抗组蛋白H1抗体染色时，亮点位于轴突和细胞核的横截面，分别。证实了可以成功地鉴定基底上的单个神经元中的靶向蛋白质。免疫GNP染色FIB/SEM观察的发展将为神经元在各种基质结构上的稳定生长提供重要信息，因为细胞内蛋白质的重新分布激活了基底上轴突的伸长和转动。
