In this note we present a combined Ki-67 and Feulgen stain for morphometric determination of the Ki-67 labelling index. The immunohistochemical part of this double staining technique is based on the alkaline-phosphatase-anti-alkaline-phosphatase (APAAP) method, visualizing the enzyme activity by the nitro-blue-tetrazolium chloride (NBT)/bromo-chloro-3-indolyl-phosphate (BCIP) technique. The NBT/BCIP complex resists the hydrolytic activity of the Feulgen stain. The staining method presented allows semi-automatic determination of both the total nucleus-area as well as the Ki-67 positive nucleus-area using a morphometric computer system. The Ki-67 labelling index thus achieved is based on the relative nuclear area of Ki-67 positive nuclei and is clearly more precise and efficient than the counting method using an ocular grid.
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Kolles, H., Förderer, W., Bock, R. et al. Combined Ki-67 and feulgen stain for morphometric determination of the Ki-67 labelling index. Histochemistry 100, 293–296 (1993). https://doi.org/10.1007/BF00270049
- Enzyme Activity
- Computer System
- Hydrolytic Activity
- Staining Method