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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >A novel, Golgi-Cox-based fluorescent staining method for visualizing full-length processes in primary rat neurons
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A novel, Golgi-Cox-based fluorescent staining method for visualizing full-length processes in primary rat neurons

机译：一种基于高尔基-科克斯（Golgi-Cox）的新型荧光染色方法，用于可视化原代大鼠神经元的全长过程

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摘要

The Golgi method, a well-known method used for staining whole dendrites and axonal trees of neurons, has been used widely for studying dendritic growth in vivo. Although detailed structural examination of neurons and their processes stained by the Golgi method has elucidated the complicated neuronal circuit, application of the method in cultured neurons has been unsuccessful to date. Here, we report the development of a stable, highly sensitive Golgi-Cox method that allows visualization of full-length processes, including the dendritic spines and the growth cones, of cultured rat neurons. This modified staining method requires: (1) rat cultured neurons fixed with a mixture containing 4% paraformaldehyde and 12.5% glutaraldehyde before impregnation with mercury; (2) rapid freezing of the fixed neurons using dry ice; and (3) immersion of the fixed neurons in Alexa Fluor 488-conjugated goat anti-rabbit immunoglobulin-G antibody (Molecular Probes, Eugene, OR) for visualization after impregnation.
机译：高尔基法是用于对神经元的整个树突和轴突树染色的公知方法，已被广泛用于研究体内树突的生长。尽管通过高尔基体法对神经元及其过程进行了详细的结构检查已阐明了复杂的神经元回路，但迄今为止该方法在培养的神经元中的应用仍未成功。在这里，我们报告了稳定，高度敏感的高尔基-柯克斯方法的发展，该方法允许可视化培养大鼠神经元的全长过程，包括树突棘和生长锥。这种改良的染色方法要求：（1）在用汞浸渍之前，将大鼠培养的神经元用含4％多聚甲醛和12.5％戊二醛的混合物固定。 （2）用干冰快速冷冻固定的神经元; （3）将固定的神经元浸入Alexa Fluor 488偶联的山羊抗兔免疫球蛋白G抗体（Molecular Probes，Eugene，OR）中，以便在浸渍后进行观察。

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