Puchtler H, Sweat Waldrop F, Meloan S N
Histochemistry. 1983;77(4):431-45. doi: 10.1007/BF00495799.
The fluorescent brightening agent Phorwhite (Blankophor) BBU imparts intense selective fluorescence to amyloid, but this modern reagent is no longer readily available on the biological dye market. Conventional Thioflavine S and T stains require differentiation and are not specific. To improve selectivity, direct and cationic thiazole dyes were substituted in the alkaline Congo Red and the Phorwhite BBU procedure. With the former technic Diphenyl Brilliant Yellow 8G, Clayton Yellow, Thiazol Yellow, Thioflavine T and Seto Flavine T imparted strong to intense selective fluorescence to amyloid. Under the conditions of the Phorwhite BBU reaction these dyes were suitable only for formalin-fixed amyloid. Several thiazole dyes did not fluoresce. Fluorescence is a function of the molecular orbital system, the thiazole rings per se cannot induce fluorescence. Paper chromatograms indicated two or more fractions in the dyes studied. Different samples of the same dye can vary significantly in their staining and fluorescence properties. This heterogeneity is inherent in the mode of synthesis. In some cases the cationic thiazole dyes rendered certain amyloid deposits, e.g. in vessel walls, intensely fluorescent; other amyloid deposits in the same sections showed only weak fluorescence. Further studies are required to correlate these peculiar patterns with immunological data on amyloid types.
荧光增白剂Phorwhite（Blankophor）BBU可使淀粉样蛋白产生强烈的选择性荧光，但这种现代试剂在生物染料市场上已不再容易获得。传统的硫黄素S和T染色需要进行分化且不具有特异性。为了提高选择性，在碱性刚果红和Phorwhite BBU方法中用直接阳离子噻唑染料进行了替代。采用前一种技术，二苯基亮黄8G、克莱顿黄、噻唑黄、硫黄素T和濑户黄素T可使淀粉样蛋白产生强至强烈的选择性荧光。在Phorwhite BBU反应条件下，这些染料仅适用于福尔马林固定的淀粉样蛋白。几种噻唑染料不产生荧光。荧光是分子轨道系统的一种功能，噻唑环本身不能诱导荧光。纸色谱表明所研究的染料中有两个或更多的组分。同一染料的不同样品在染色和荧光特性上可能有很大差异。这种异质性是合成方式所固有的。在某些情况下，阳离子噻唑染料可使某些淀粉样沉积物（如血管壁中的沉积物）产生强烈荧光；同一切片中的其他淀粉样沉积物仅显示微弱荧光。需要进一步研究以将这些特殊模式与关于淀粉样蛋白类型的免疫学数据相关联。
