Bettinger C, Zimmermann H W
Institut für Physikalische Chemie, Universität Freiburg, Federal Republic of Germany.
Histochemistry. 1991;96(3):215-28. doi: 10.1007/BF00271540.
The absorption spectra of hematein-aluminium solutions have been recorded at various concentrations and pH values; the solutions were prepared using analytically pure hematein and potassium alum as aluminium source. In aqueous solution, four different hematein-aluminium complexes could be distinguished by absorption spectroscopy. In weakly acidic media we observed the violet 1:1 and 1:2 complexes HmAl (VII) and HmAl2(3) (VIII), and in strongly acidic solution the red 1:1 complex HmAl2 (IX). Whereas, in weakly alkaline solution the blue 1:1 complex HmAl0 (X) was detected. By change of the pH value the complexes were mutual interconverted. The dye complexes were characterized by their absorption spectra and molar extinction coefficients. We have stained HeLa cells with the complex solutions under different experimental conditions. In all cases the nuclear staining was intense whereas the staining of the cytoplasm was weak. The microspectra of the stained nuclei were recorded and compared with the absorption spectra of the complexes in solution. Thus it was possible to identify the bound dye species. After staining in acidic media, the cells were red to red-violet depending on the reaction conditions. The three cationic dye species VII, VIII, and IX were bound in varying amounts. After blueing in weakly acidic media or in water, only the violet dye complex VII was detected whereas, after blueing in weakly alkaline media, only the blue complex X has been observed. Enzymatic digestion experiments have shown that the dye complexes in the nuclei were bound to DNA while those in the cytoplasm and nucleoli were bound to RNA. The binding between the dye complexes and the nucleic acids is discussed.
已记录了苏木精 - 铝溶液在不同浓度和pH值下的吸收光谱；溶液使用分析纯苏木精和硫酸铝钾作为铝源制备。在水溶液中，通过吸收光谱可区分出四种不同的苏木精 - 铝配合物。在弱酸性介质中，我们观察到紫色的1:1和1:2配合物HmAl（VII）和HmAl2(3)（VIII），在强酸性溶液中观察到红色的1:1配合物HmAl2（IX）。而在弱碱性溶液中检测到蓝色的1:1配合物HmAl0（X）。通过改变pH值，这些配合物可相互转化。这些染料配合物通过其吸收光谱和摩尔消光系数进行表征。我们在不同实验条件下用配合物溶液对HeLa细胞进行了染色。在所有情况下，细胞核染色强烈，而细胞质染色较弱。记录了染色细胞核的显微光谱，并与溶液中配合物的吸收光谱进行比较。因此能够鉴定出结合的染料种类。在酸性介质中染色后，根据反应条件细胞呈红色至红紫色。三种阳离子染料种类VII、VIII和IX以不同量结合。在弱酸性介质或水中进行返蓝后，仅检测到紫色染料配合物VII，而在弱碱性介质中返蓝后，仅观察到蓝色配合物X。酶消化实验表明，细胞核中的染料配合物与DNA结合，而细胞质和核仁中的染料配合物与RNA结合。讨论了染料配合物与核酸之间的结合。
