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Combined Ki-67 and feulgen stain for morphometric determination of the Ki-67 labelling index

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Abstract

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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References

  1. Böck P (1989) ROMEIS — Mikroskopische Technik. Urban and Schwarzenberg, Munich, pp 352–353

  2. Cattoretti G, Becker MHG, Key G, Duchrow M, Schlüter C, Galle J, Gerdes J (1992) Monoclonal antibodies against recombinant parts of the Ki-67 antigen (MIB 1 and MIB 3) detect proliferating cells in microwave-processed formalin-fixed paraffin sections. J Pathol 168:357–363

  3. Chatelain R, Willms A, Biesterfeld S, Auffermann W, Böcking A (1989) Automated Feulgen staining with a temperature-controlled staining machine. Anal Quant Cytol Histol 11:211–217

  4. De Tomasi JA (1936) Improving the technic of the Feulgen stain. Stain Techol 11:137–144

  5. Feulgen F, Rossenbeck H (1924) Mikroskopisch-chemischer Nachweis einer Nucleinsäure vom Typ der Thymonucleinsäure und die darauf beruhende elektive Färbung von Zellkernen in mikroskopischen Präparaten. Hoppe Seylers Z Physiol Chem 135:203–248

  6. Gerdes J, Schwab U, Lemke H, Stein H (1983) Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. Int J Cancer 31:13–20

  7. Gerdes J, Becker MHG, Key G, Cattoretti G (1992) Immunohistological detection of tumor growth fraction (Ki-67 antigen) in formalin-fixed and routinely processed tissues. J Pathol 168:85–87

  8. Horwitz JP, Chua J, Noel M, Donatti JT, Freisler J (1966) Substrates for cytochemical demonstration of enzyme activity. J Med Chem 9:447

  9. Kiefer G, Kiefer R, Moore GW, Salm R, Sandritter W (1974) Nuclear images of cells in different functional states. J Histochem Cytochem 22:569–576

  10. Martin H, Voss K, Hufnagl P, Frölich K (1984) Automated image analysis of gliomas. An objective and reproducible method for tumor grading. Acta Neuropathol 63:160–169

  11. Mikel UV, Becker RL (1991) A comparative study of quantitative stains for DNA in image cytometry. Anal Quant Cytol Histol 13:253–260

  12. Sawhney N, Hall PA (1992) Ki-67 — structure, function, and new antibodies. J Pathol 168:161–162

  13. Schröder R, Bien K, Kott R, Meyers I, Vössing R (1991) The relationship between Ki-67 labelling and mitotic index in gliomas and meningiomas: demonstration of the variability of the intermitotic cycle time. Acta Neuropathol 82:389–394

  14. Shi S-R, Key ME, Kalra KL (1991) Antigen retrieval in formalin-fixed, paraffin-embedded tissues: An enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections. J Histochem Cytochem 39:741–748

  15. Van Dierendonck JH, Wijsman JH, Keijzer R, Van de Velde CJH, Cornelisse CJ (1991) Cell-cycle-related staining patterns of anti-proliferating cell nuclear antigen monoclonal antibodies. Comparison with BrdUrd labeling and Ki-67 staining. Am J Pathol 138:1165–1172

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Correspondence to Harry Kolles.

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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

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Keywords

  • Chloride
  • Enzyme Activity
  • Computer System
  • Hydrolytic Activity
  • Staining Method