Joseph Sabitha, Werner Hauke B, Stegmüller Judith
Department of Neurology, RWTH Aachen University Hospital, Pauwelsstrasse 30, 52074 Aachen, Germany.
Department of Neurogenetics, Max Planck Institute of Experimental Medicine, Hermann Rein Strasse 3, 37075 Göttingen.
Bio Protoc. 2019 Nov 20;9(22):e3436. doi: 10.21769/BioProtoc.3436.
In the nervous system of vertebrates, nerve impulse propagation is accelerated by the ensheathment of neuronal axons with myelin. Myelin sheaths are molecularly specialized, lipid-rich plasma membrane extensions of Schwann cells in the peripheral nervous system and oligodendrocytes in the central nervous system (CNS). To visualize myelinated nerve fibers and to allow for the morphological analyses of myelin in the brain and the spinal cord, an efficient method for silver impregnation of myelin has originally been developed by Ferenc Gallyas in 1979, referred to as . Gallyas' method is based on the agyrophilic characteristic of myelin to form and bind silver particles, while this process is suppressed in tissues other than myelin. The silver particles are finally enhanced in a developing step ("physical developer"). The main advantage of this method is that it efficiently visualizes both large myelinated fiber tracts and individual myelinated axons. Here we provide our laboratory protocol that is suitable for paraffin embedded sections and the use of light microscopy based on Gallyas' original protocol and subsequent modifications by Pistorio and colleagues.
在脊椎动物的神经系统中，神经元轴突被髓鞘包裹可加速神经冲动的传播。髓鞘是外周神经系统中施万细胞和中枢神经系统（CNS）中少突胶质细胞富含脂质的分子特异性质膜延伸物。为了可视化有髓神经纤维并对脑和脊髓中的髓鞘进行形态学分析，费伦茨·加利亚什于1979年最初开发了一种有效的髓鞘银浸染方法，称为加利亚什法。加利亚什法基于髓鞘形成并结合银颗粒的嗜银特性，而在髓鞘以外的组织中该过程受到抑制。银颗粒最终在显影步骤（“物理显影剂”）中得到增强。该方法的主要优点是它能有效地可视化大型有髓纤维束和单个有髓轴突。在此，我们基于加利亚什的原始方案以及皮斯托里奥及其同事随后的修改，提供适用于石蜡包埋切片和光学显微镜使用的实验室方案。
