Template:Immunohistochemistry evaluation of breast cancer

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Ki-67 index

Ki-67 in an invasive breast cancer: cancer nuclei are stained (brown). There is tumor cell positivity in 70% of the cells (Ki-67 labelling index = 70%).

For Ki-67 index, there are two main methods, a faster "hot spot" method and the more comprehensive IKWG global average:

Hot spot:

Hot spots are areas in which Ki-67 staining is particularly higher relative to the adjacent tumor areas.[1] Usually, the invasive edge of a tumor is a hot spot.[1] When a tumor had several hot spots, the “hottest” spot is selected.[1] Aim to count at least 500 cells in each case, but this is not always possible in cases with low tumor cell density and small tumor size.[1] Also aim to include at least three high-power (×40 objective) fields.[2]

IKWG global average:[3]
  1. Before first use, access the IKWG website (https://www.ki67inbreastcancerwg.org/) and complete the Ki67 calibration exercise
  2. From Tools, link to the Online scoring app (or download and install the Ki67 counting app) and use the global method
  3. Using a regular light microscope, review the Ki67-stained breast cancer slide and input estimates of the percent area with negligible, low, medium, or high Ki67 index
  4. Score 100 nuclei negative or positive in each field type (as directed by the app)
  5. Record “Weighted global score” output as the Ki67 index for that slide

Regardless of method:[2]

  • If a comparisons must be made between core biopsies and sections from an excision, evaluation of the latter should be across the whole tumor.
  • Only nuclear staining counts. Staining intensity of a positive nucleus is not relevant.


Score[4] Status[4]
0 to 1+ HER2 negative
(not present)
2+ Borderline
3+ HER2 positive


Main page


  1. 1.0 1.1 1.2 1.3 Coleman, William B.; Jang, Min Hye; Kim, Hyun Jung; Chung, Yul Ri; Lee, Yangkyu; Park, So Yeon (2017). "A comparison of Ki-67 counting methods in luminal Breast Cancer: The Average Method vs. the Hot Spot Method ". PLOS ONE 12 (2): e0172031. doi:10.1371/journal.pone.0172031. ISSN 1932-6203. 
  2. 2.0 2.1 Dowsett, M.; Nielsen, T. O.; A'Hern, R.; Bartlett, J.; Coombes, R. C.; Cuzick, J.; Ellis, M.; Henry, N. L.; et al. (2011). "Assessment of Ki67 in Breast Cancer: Recommendations from the International Ki67 in Breast Cancer Working Group ". JNCI Journal of the National Cancer Institute 103 (22): 1656–1664. doi:10.1093/jnci/djr393. ISSN 0027-8874. 
  3. Nielsen, Torsten O; Leung, Samuel C. Y; Rimm, David L; Dodson, Andrew; Acs, Balazs; Badve, Sunil; Denkert, Carsten; Ellis, Matthew J; et al. (2021). "Assessment of Ki67 in Breast Cancer: Updated Recommendations From the International Ki67 in Breast Cancer Working Group ". JNCI: Journal of the National Cancer Institute 113 (7): 808–819. doi:10.1093/jnci/djaa201. ISSN 0027-8874. 
  4. 4.0 4.1 . IHC Tests (ImmunoHistoChemistry). Breastcancer.org. Retrieved on 2019-10-04. Last modified on October 23, 2015