Castejón H V, Castejón O J, Viloria M E
Acta Histochem. 1976;56(2):285-300. doi: 10.1016/S0065-1281(76)80116-X.
Alcian blue is a cationic dye which has been used in the histochemical field for the demonstration of polyanions especially carboxylated and sulphated. The results obtained in neurons when this dye was applied to human and mouse cerebral cortex and studied with the electron microscope are the object of the present report. The CNS of normal adult mice was fixed by vascular perfusion with 2% glutaraldehyde-0.1 M sodium cacodylate-0.1 M sucrose at pH = 6.8 followed by the same fixative with the addition of 0.5% alcian blue. After perfusion, brain cortex was taken out, sectioned into small blocks and immersed in a fresh similar mixture and subsequently in OSO4. Blocks were dehydrated and embedded in araldite. Ultrathin sections were doubly stained with uranyl and lead salts. Human brain cortex taken from patients with cerebral edema was fixed by immersion with 6.5% glutaraldehyde-0.1 M sodium phosphate, pH = 7.4 followed by embedding in warm agar and sectioning in slices of 30 mum thickness which were impregnated by immersion in a mixture of 1% alcian blue-acetate buffer-3% glutaraldehyde at pH = 3.5 for 9 to 15 h at 4 degrees C and subsequently immersed in 1% buffered OSO4-0.1 M sucrose, pH = 7.4 for 2 h at 4 degrees S. Sections were dehydrated and embedded in araldite. Ultrathin sections were doubly stained by uranyl and lead salts. We have denominated the complete procedure in both instances GABOUL technique. The submicroscopic study of both tissues, at nerve cells, revealed the presence of an electron dense homogeneous substance thoroughly dispersed at the hyaloplasmic matrix of perikarya, processes and even synaptic endings. This substance was more evident around free and attached ribosomes, GOLGI apparatus, complex vesicles, dense bodies, microtubules, subsurface cisternae and synaptic vesicles. Canaliculi of endoplasmic reticulum and even the perinuclear cistern also showed a moderate content. It is suggested that this electron dense substance, being alcianophilic, has a polyanionic character and thus may partially correspond to acid polysaccharides since these compounds have already been previously confirmed in such neurons by biochemical and light microscope histochemical techniques.
阿尔辛蓝是一种阳离子染料，已在组织化学领域用于显示多阴离子，尤其是羧化和硫酸化的多阴离子。将这种染料应用于人和小鼠大脑皮层并用电子显微镜研究时，在神经元中获得的结果是本报告的主题。正常成年小鼠的中枢神经系统通过血管灌注用pH = 6.8的2%戊二醛-0.1M二甲胂酸钠-0.1M蔗糖固定，随后用添加了0.5%阿尔辛蓝的相同固定剂固定。灌注后，取出大脑皮层，切成小块，浸入新鲜的类似混合物中，随后浸入四氧化锇中。小块脱水并包埋在环氧树脂中。超薄切片用铀盐和铅盐进行双重染色。取自脑水肿患者的人脑皮层通过浸入6.5%戊二醛-0.1M磷酸钠（pH = 7.4）固定，随后包埋在温热的琼脂中，并切成30μm厚的切片，将其浸入pH = 3.5的1%阿尔辛蓝-醋酸缓冲液-3%戊二醛混合物中在4℃下浸渍9至15小时，随后浸入1%缓冲四氧化锇-0.1M蔗糖（pH = 7.4）中在4℃下浸渍2小时。切片脱水并包埋在环氧树脂中。超薄切片用铀盐和铅盐进行双重染色。我们将这两种情况下的完整程序命名为加布尔技术。对这两种组织在神经细胞处的亚显微研究表明，在核周体、突起甚至突触末梢的透明质基质中存在一种电子致密的均匀物质，且分布均匀。这种物质在游离和附着的核糖体、高尔基体、复合囊泡、致密体、微管、表面下池和突触小泡周围更为明显。内质网的小管甚至核周池也显示出适度的含量。有人认为，这种电子致密物质具有亲阿尔辛蓝性，具有多阴离子特性，因此可能部分对应于酸性多糖，因为这些化合物先前已通过生化和光学显微镜组织化学技术在这类神经元中得到证实。
