Tas J, van der Ploeg M, James J
Acta Histochem Suppl. 1979;20:69-73.
After staining with Naphthol Yellow S (NYS) at optimal conditions of pH (2.8), the protein content of rat liver cells isolated by means of a collagenase perfusion technique was found to be cytophotometrically immeasurable, because of too high local dye absorbances. In order to lower the absorption values, techniques to flatten the cells, off-peak measurements and NYS staining at non-optimal pH levels were applied. With polyacrylamide model films incorporated with albumin, the reliability of off-peak measurements and the quantitative aspects of the modified protein staining procedures have been investigated. It was found that NYS is a quantitative protein stain not only at pH 2.8, but also at pH 2.0, 3.5 and 4.0 respectively. Off-peak measurements can also produce quantitative results. In cases of a combined Feulgen-NYS staining, the Feulgen-DNA values were not significantly influenced by any of the NYS staining procedures.
在pH(2.8)的最佳条件下用萘酚黄S(NYS)染色后,发现通过胶原酶灌注技术分离的大鼠肝细胞的蛋白质含量由于局部染料吸光度太高而无法用细胞光度法测量。为了降低吸光度值,采用了使细胞变平的技术、非峰值测量以及在非最佳pH水平下进行NYS染色。对于掺入白蛋白的聚丙烯酰胺模型膜,研究了非峰值测量的可靠性以及改良蛋白质染色程序的定量方面。结果发现,NYS不仅在pH 2.8时是一种定量蛋白质染色剂,而且分别在pH 2.0、3.5和4.0时也是。非峰值测量也可以产生定量结果。在福尔根-NYS联合染色的情况下,福尔根-DNA值不受任何NYS染色程序的显著影响。