Roe R, Corfield A P, Williamson R C
University Department of Surgery, Bristol Royal Infirmary, UK.
Histochem J. 1989 Apr;21(4):216-22. doi: 10.1007/BF01747523.
Two new histochemical procedures for detecting sulphated and non-sulphated sialomucin in colonic mucosa were assessed: the saponification-Alcian Blue pH 1-periodic acid-phenylhydrazine-Schiff method (KOH-AB pH 1-PAPS) and the mild periodic acid modification of this (KOH-AB pH 1-mPAS). Using normal colonic mucosa obtained from 11 non-cancer patients, the mPAS and PAPS techniques were tested for specificity and reproducibility for staining sialic acid, either alone or in combination with Alcian Blue. A spectrophotometric method was devised to quantify the uptake of both Schiff and Alcian Blue stain by sections. At low temperature and pH 5.5, the mPAS procedure had improved specificity over the PAPS procedure, and after saponification it could be used to stain O-acetyl-substituted sialic acid. When used in combination with Alcian Blue at pH 1, however, underestimation of the sialic acid content occurred owing to interference between Alcian Blue and Schiff dyes. Interference was even greater with KOH-AB pH 1-PAPS procedure for both sialic acid and sulphate components. We conclude that caution must be exercised in interpretation of the staining results obtained with these new combination methods and that more accurate information on the sialic acid and sulphate content of colonic mucin is obtained by staining serial sections with the mPAS technique and Alcian Blue pH 1 alone.
皂化-阿尔辛蓝pH 1-高碘酸-苯肼-席夫法（KOH-AB pH 1-PAPS）及其温和高碘酸改良法（KOH-AB pH 1-mPAS）。使用从11名非癌症患者获取的正常结肠黏膜，对mPAS和PAPS技术单独或与阿尔辛蓝联合染色唾液酸的特异性和可重复性进行了测试。设计了一种分光光度法来量化切片对席夫和阿尔辛蓝染料的摄取量。在低温和pH 5.5条件下，mPAS程序比PAPS程序具有更高的特异性，皂化后可用于染色O-乙酰基取代的唾液酸。然而，当与pH 1的阿尔辛蓝联合使用时，由于阿尔辛蓝和席夫染料之间的干扰，会低估唾液酸含量。对于唾液酸和硫酸盐成分，KOH-AB pH 1-PAPS程序的干扰更大。我们得出结论，在解释这些新联合方法获得的染色结果时必须谨慎，并且通过单独用mPAS技术和pH 1的阿尔辛蓝对连续切片进行染色，可以获得关于结肠黏蛋白中唾液酸和硫酸盐含量更准确的信息。
