Approaches utilizing multiple analysis techniques on a single sample are highly desirable in research, especially to reduce the number of animals and obtain the maximum information. Golgi-Cox staining is a widely used method for characterizing axon and dendritic morphology and several attempts to combine this technique with immunofluorescence and transmission electron microscopy have been proposed. With few exceptions, most of the protocols were characterized by a high degree of complexity and low reproducibility. Here we show a simplified procedure of perfusion, fixation and staining of brain tissues that allows Golgi-Cox staining, immunofluorescence and transmission electron microscopy in the same sample, to obtain high-quality images with a low-cost procedure. The main novelty in this protocol is the possibility of performing Golgi-Cox staining after the perfusion and post-fixation of brain tissue with a buffered solution containing, not only formaldehyde, but also glutaraldehyde. This renders the tissue suitable for electron microscopy, but it is also compatible with immunofluorescence staining. This combined protocol can be used in most neuroscience laboratories as it does not require special equipment and skills. This protocol will be useful in a broad range of neuroscience topics to study morphological changes during brain development and plasticity in physiological and pathological conditions.

在研究中非常需要在单个样本上使用多种分析技术的方法，特别是为了减少动物数量并获得最大信息。高尔基-考克斯染色是一种广泛使用的表征轴突和树突形态的方法，并且已经提出了将这种技术与免疫荧光和透射电子显微镜相结合的几种尝试。除了少数例外，大多数协议的特点是高度复杂和低可重复性。在这里，我们展示了一种简化的脑组织灌注、固定和染色程序，允许在同一样本中进行高尔基-考克斯染色、免疫荧光和透射电子显微镜检查，从而以低成本的程序获得高质量的图像。该协议的主要新颖之处在于有可能在脑组织灌注和固定后进行高尔基-考克斯染色，缓冲溶液不仅含有甲醛，还含有戊二醛。这使得组织适用于电子显微镜，但它也与免疫荧光染色兼容。这种组合协议可用于大多数神经科学实验室，因为它不需要特殊设备和技能。该协议将在广泛的神经科学主题中有用，以研究生理和病理条件下大脑发育和可塑性的形态变化。但它也与免疫荧光染色兼容。这种组合协议可用于大多数神经科学实验室，因为它不需要特殊设备和技能。该协议将在广泛的神经科学主题中有用，以研究生理和病理条件下大脑发育和可塑性的形态变化。但它也与免疫荧光染色兼容。这种组合协议可用于大多数神经科学实验室，因为它不需要特殊设备和技能。该协议将在广泛的神经科学主题中有用，以研究生理和病理条件下大脑发育和可塑性的形态变化。