Tolivia J, Navarro A, Tolivia D
Departamento de Morfología y Biología Celular, Facultad de Biología y Medicina, Universidad de Oviedo, Spain.
Histochemistry. 1994 Aug;102(2):101-4. doi: 10.1007/BF00269013.
A differential staining method is described of myelinated fibres and nerve cell bodies applicable to sections of mammalian, including human, central nervous system specimens embedded in paraffin wax. Experimental and human necropsy material fixed in acetic paraformaldehyde in phosphate buffer was used. Sections of 15-20 microns in thickness were obtained, attached to slides, deparaffinized and hydrated. After hydration, sections are oxidized (30 s) in 2% potassium permanganate, bleached (1 min) in 5% oxalic acid and rinsed in distilled water. Staining is for 2-5 h in the following solution: 0.06% thionin, 1% formaldehyde, 10% acetic acid in distilled water. Sections are subsequently washed in distilled water, dehydrated through 96% and absolute ethanol, cleared in eucalyptol and mounted in Eukitt. Using the method described in the present paper, a differential coloration of myelin and neurons is obtained. Myelinated fibres appear red, whereas nerve cell bodies and glial nuclei are stained blue. This procedure provides a high contrast between myelin and cells suitable for observation and photography of sections. Simultaneous and differential coloration of both myelin and cells is easily and directly obtained with constant and homogeneous results.
本文描述了一种适用于石蜡包埋的哺乳动物（包括人类）中枢神经系统标本切片的有髓纤维和神经细胞体的鉴别染色方法。使用了固定于磷酸盐缓冲液中的乙酸多聚甲醛中的实验材料和人体尸检材料。制备厚度为15 - 20微米的切片，贴于载玻片上，脱蜡并水化。水化后，切片在2%高锰酸钾中氧化（30秒），在5%草酸中漂白（1分钟），然后用蒸馏水冲洗。在以下溶液中染色2 - 5小时：0.06%硫堇、1%甲醛、蒸馏水中的10%乙酸。随后切片用蒸馏水洗涤，经96%乙醇和无水乙醇脱水，用桉叶油透明，并用尤基特胶封固。使用本文所述方法，可使髓磷脂和神经元呈现不同颜色。有髓纤维呈红色，而神经细胞体和神经胶质细胞核染成蓝色。此方法能使髓磷脂与细胞之间形成高对比度，适合对切片进行观察和拍照。可轻松直接地同时实现髓磷脂和细胞的鉴别染色，结果恒定且均匀。
