Gossrau R
Histochemistry. 1978 Dec 1;58(3):203-18. doi: 10.1007/BF00495720.
Using freeze-dried or sections from fresh-frozen or aldehyde-fixed material nitro BT (NBT), tetranito BT (TNBT), distyryl nitro BT (DS-NBT), thiocarbamyl nitro BT (TC-NBT) or benzothiazolylstyrylphthalhydrazidyl tetrazolium chloride (BSPT) were tested as auxiliary reagents for the localization of glycosidases, phosphatases and non-specific esterases with indoxyl substrates in rat tissues. By means of NBT or TNBT as a tetrazolium salt acid beta-D-galactosidase, beta-D-N-acetylglucosaminidase, acid phosphatase, neuraminidase and non-specific esterase can only be localized at the cellular level; a more precise localization is possible for lactase-beta-D-glucosidase in the intestinal brush border, and the best results are obtained in the demonstration of alkaline phosphatase; among all methods described previously the tetrazolium procedure with TNBT is the method of choice for the light microscopic localization of this enzyme. Reverse data are observed with BSPT as a tetrazolium salt; then, all acid and neutral hydrolases can be exactly localized in lysosomes, secretion granules, cytoplasm and/or microvilli of many cells and tissues provided BSPT-formazan is stabilized by osmification. Furthermore, this procedure enables the reliable ultracytochemical demonstration of these enzymes. However, in the case of alkaline phosphatase only sites with high enzyme activity reveal a positive reaction. -DS- and TC-NBT are inferior to NBT, TNBT or BSPT.
使用来自冻干材料、新鲜冷冻材料或醛固定材料的切片，对硝基蓝四唑（NBT）、四硝基蓝四唑（TNBT）、二苯乙烯基硝基蓝四唑（DS-NBT）、硫代氨基甲酰硝基蓝四唑（TC-NBT）或苯并噻唑基苯乙烯基邻苯二甲酰肼基氯化四唑（BSPT）作为辅助试剂，用于在大鼠组织中用吲哚酚底物定位糖苷酶、磷酸酶和非特异性酯酶。借助NBT或TNBT作为四唑盐，酸性β-D-半乳糖苷酶、β-D-N-乙酰氨基葡萄糖苷酶、酸性磷酸酶、神经氨酸酶和非特异性酯酶只能在细胞水平上定位；乳糖酶-β-D-葡萄糖苷酶在肠刷状缘可以进行更精确的定位，而在碱性磷酸酶的显示中获得了最佳结果；在先前描述的所有方法中，用TNBT的四唑法是该酶光镜定位的首选方法。以BSPT作为四唑盐观察到相反的数据；然后，只要通过锇化使BSPT-甲臜稳定，所有酸性和中性水解酶都可以在许多细胞和组织的溶酶体、分泌颗粒、细胞质和/或微绒毛中准确定位。此外，该方法能够可靠地对这些酶进行超微细胞化学显示。然而，对于碱性磷酸酶，只有酶活性高的部位显示阳性反应。DS-NBT和TC-NBT不如NBT、TNBT或BSPT。
