Araki T, Chikamori K, Sasaki K, Kawata S, Minami S, Yamada M
Histochemistry. 1987;86(6):567-72. doi: 10.1007/BF00489548.
A component-spectroanalysis technique was used to study the multicolor properties of histochemically stained tissue sections. We developed a method that makes it possible to obtain separately both the spectral patterns and spatial distributions of different color components in tissue sections. To illustrate the application of this technique, we examined the extinction spectrum of reduced nitroblue tetrazolium (NBT), which is used for the detection of dehydrogenase activity. Upon the reduction of NBT, mono- and diformazans are formed, and these exhibit over-lapping extinction spectra. When succinate dehydrogenase (SDH) activity in rat liver lobules was examined using NBT, monoformazan was found to be present at higher concentrations than diformazan and to have a uniform distribution, whereas the concentration of diformazan increased with a steep gradient between the center and periphery of lobules. In rat skeletal muscle fibers, diformazan was present at higher concentrations than monoformazan. The level of SDH activity was topographically represented by the hydrogen concentration calculated from the concentrations of the two formazans. This method is effective for separating multiple components such as mono- and diformazans in histochemical reactions.
采用成分光谱分析技术研究组织化学染色组织切片的多色特性。我们开发了一种方法，该方法能够分别获取组织切片中不同颜色成分的光谱模式和空间分布。为说明该技术的应用，我们检测了用于检测脱氢酶活性的还原型硝基蓝四氮唑（NBT）的消光光谱。NBT还原后会形成单甲臜和二甲臜，且它们呈现出重叠的消光光谱。当使用NBT检测大鼠肝小叶中的琥珀酸脱氢酶（SDH）活性时，发现单甲臜的浓度高于二甲臜且分布均匀，而二甲臜的浓度在小叶中心和周边之间呈陡峭梯度增加。在大鼠骨骼肌纤维中，二甲臜的浓度高于单甲臜。SDH活性水平由根据两种甲臜的浓度计算出的氢浓度进行拓扑表示。该方法对于在组织化学反应中分离单甲臜和二甲臜等多种成分是有效的。
