Oud P S, Henderik J B, Huysmans A C, Pahlplatz M M, Hermkens H G, Tas J, James J, Vooijs G P
Histochemistry. 1984;80(1):49-57. doi: 10.1007/BF00492771.
The protein dyes Light Green and Orange II were studied separately and in combination with the Feulgen-Pararosanilin(SO2) and -Thionin(SO2) method for the simultaneous determination of DNA and protein. - With polyacrylamide modelfilms the pH dependency, specificity and stoichiometry of Light Green and Orange II have been investigated. The results of both staining methods with different biological objects have been compared. - In addition, the Feulgen-Thionin(SO2) method was studied with model films with respect to its specificity and stoichiometry. In biological objects it has been compared with the Feulgen-Pararosanilin(SO2) method. - When combining the Light Green staining with the Feulgen-Pararosanilin(SO2) procedure and the Orange II staining with Feulgen-Thionin(SO2), both Feulgen-DNA stainings, which were first applied, proved to be unaffected by the following protein staining procedure. When the Feulgen procedure was carried out without the dye, followed by Light Green staining, the latter became reduced when a sulfite water rinse was included but was unaffected when a running tap water rinse was used. In the case of the Orange II staining a serious reduction in dye binding capacity was found in both situations. - When the Feulgen-Pararosanilin(SO2) Light Green procedure was carried out on isolated nuclei with all dyes present, a decrease of protein dye binding was observed, similar to that found with the well-known Feulgen-Pararosanilin(SO2) Naphthol Yellow S combination. It is concluded that in spite of this reduction the latter two combinations can be used for the cytophotometric analysis of DNA and protein in the same object.
分别研究了蛋白质染料亮绿和橙黄II，并将它们与福尔根-副蔷薇苯胺（SO2）和-硫堇（SO2）方法结合，用于同时测定DNA和蛋白质。- 使用聚丙烯酰胺模型膜研究了亮绿和橙黄II的pH依赖性、特异性和化学计量关系。比较了两种染色方法对不同生物样品的染色结果。- 此外，还使用模型膜研究了福尔根-硫堇（SO2）方法的特异性和化学计量关系。在生物样品中，将其与福尔根-副蔷薇苯胺（SO2）方法进行了比较。- 当将亮绿染色与福尔根-副蔷薇苯胺（SO2）程序相结合，以及将橙黄II染色与福尔根-硫堇（SO2）相结合时，首先进行的两种福尔根-DNA染色均未受随后的蛋白质染色程序影响。当在没有染料的情况下进行福尔根程序，随后进行亮绿染色时，如果用亚硫酸盐水溶液冲洗，亮绿会褪色，但用自来水冲洗则不受影响。对于橙黄II染色，在这两种情况下均发现染料结合能力严重下降。- 当在所有染料都存在的情况下，对分离的细胞核进行福尔根-副蔷薇苯胺（SO2）亮绿程序时，观察到蛋白质染料结合减少，这与著名的福尔根-副蔷薇苯胺（SO2）萘酚黄S组合的情况相似。得出的结论是，尽管存在这种减少，但后两种组合仍可用于同一物体中DNA和蛋白质的细胞光度分析。
