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采用雕白粉－亚甲蓝还原液对大白鼠和家兔的迷走神经和坐骨神经进行活体染色，并采用钼酸胺与四氧化锇固定，纯乙醇脱水的方法，制作组织切片和超薄切片，进行光镜和电镜研究。光镜标本显示，细神经均匀染色，而粗神经仅浅层染色，有髓神经纤维的轴突深染，但髓鞘不染色。电镜可见染色标本神经纤维内的微管和神经微丝，认为还原型亚甲蓝是以Ｈ２Ｂ＋形式穿透细胞膜（或轴膜），在轴突内氧化成亚甲蓝阳离子，与负电荷基团结合，使神经纤维显示蓝色。
Vivo staining of the vagus nerve and sciatic nerve in rats and white rabbits by carved white powder-methylene blue reduction solution was performed. Tissue sections and ultrathin sections were made by fixed molybdate and osmium tetroxide and pure ethanol dehydration. Mirror and electron microscopy. Light microscopy showed that the fine nerves were stained evenly, whereas the superficial nerve was only superficially dyed, the axons of myelinated fibers were deeply stained, but myelin was not stained. The microtubules and the nerve microfilaments in the nerve fibers of the stained specimens were observed by electron microscopy. The reduced methylene blue is thought to penetrate the cell membrane (or axial membrane) in the form of H2B + and be oxidized into methylene blue cations in the axons, and the negatively charged groups Combined, make nerve fibers appear blue.