Fukuda M, Nakanishi K, Böhm N, Kimura J, Harada K, Fujita S
Histochemistry. 1979 Sep;63(1):35-45. doi: 10.1007/BF00508010.
Feulgen nuclear staining with pararosanilin-SO2 was combined with the ninhydrin-Schiff technique. The aldehyde groups converted from primary amino groups are stained with an acriflavine-Schiff reaction. This results in a red nuclear fluorescence and a bright yellow cytoplasmic and nuclear fluorescence. The combined fluorescence staining facilitates cytofluorometric determination of total protein and DNA in the same cell. The ninhydrin-Schiff reaction is affected by the fixation procedure and the duration of the ninhydrin reaction. Investigations with a model system showed that proportionality between the fluorescence intensity of acriflavine and the amount of protein stained by the procedure was obtained after fixation with a fixation mixture suggested by Böhm et al. (1968) and a reaction with ninhydrin at 37 degrees C for 10 h. The ninhydrin-Schiff reaction has no effect on the fluorescence intensity of cells previously treated with pararosanilin-Feulgen staining and it is not affected itself by this previous procedure. Testing this double fluorescence staining on cytology specimens taken from patients with gastric carcinoma and uterine cervial carcinoma, cancer cells were shown to have markedly increased protein and DNA contents compared with those of normal cells.
将副品红 - SO₂ 的福尔根核染色与茚三酮 - 席夫技术相结合。由伯氨基转化而来的醛基通过吖啶黄素 - 席夫反应进行染色。这会产生红色的核荧光以及明亮的黄色细胞质和核荧光。这种联合荧光染色有助于在同一细胞中进行细胞荧光测定总蛋白和 DNA。茚三酮 - 席夫反应受固定程序和茚三酮反应持续时间的影响。用模型系统进行的研究表明，在用 Böhm 等人（1968 年）建议的固定混合物固定并在 37℃下与茚三酮反应 10 小时后，吖啶黄素的荧光强度与该程序染色的蛋白量之间呈比例关系。茚三酮 - 席夫反应对先前用副品红 - 福尔根染色处理过的细胞的荧光强度没有影响，并且它本身也不受此先前程序的影响。对取自胃癌和子宫颈癌患者的细胞学标本进行这种双重荧光染色测试时，显示癌细胞的蛋白质和 DNA 含量与正常细胞相比明显增加。
