+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Proliferation assessment in breast carcinomas using digital image analysis based on virtual Ki67/cytokeratin double staining

Proliferation assessment in breast carcinomas using digital image analysis based on virtual Ki67/cytokeratin double staining

Breast Cancer Research and Treatment 158(1): 11-19

Manual estimation of Ki67 Proliferation Index (PI) in breast carcinoma classification is labor intensive and prone to intra- and interobserver variation. Standard Digital Image Analysis (DIA) has limitations due to issues with tumor cell identification. Recently, a computer algorithm, DIA based on Virtual Double Staining (VDS), segmenting Ki67-positive and -negative tumor cells using digitally fused parallel cytokeratin (CK) and Ki67-stained slides has been introduced. In this study, we compare VDS with manual stereological counting of Ki67-positive and -negative cells and examine the impact of the physical distance of the parallel slides on the alignment of slides. TMAs, containing 140 cores of consecutively obtained breast carcinomas, were stained for CK and Ki67 using optimized staining protocols. By means of stereological principles, Ki67-positive and -negative cell profiles were counted in sampled areas and used for the estimation of PIs of the whole tissue core. The VDS principle was applied to both the same sampled areas and the whole tissue core. Additionally, five neighboring slides were stained for CK in order to examine the alignment algorithm. Correlation between manual counting and VDS in both sampled areas and whole core was almost perfect (correlation coefficients above 0.97). Bland-Altman plots did not reveal any skewness in any data ranges. There was a good agreement in alignment (>85 %) in neighboring slides, whereas agreement decreased in non-neighboring slides. VDS gave similar results compared with manual counting using stereological principles. Introduction of this method in clinical and research practice may improve accuracy and reproducibility of Ki67 PI.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 058641006

Download citation: RISBibTeXText

PMID: 27283833

DOI: 10.1007/s10549-016-3852-6

Related references

Digital image analysis of Ki67 proliferation index in breast cancer using virtual dual staining on whole tissue sections: clinical validation and inter-platform agreement. Breast Cancer Research and Treatment 169(1): 33-42, 2018

Image Analysis in Digital Pathology: Combining Automated Assessment of Ki67 Staining Quality with Calculation of Ki67 Cell Proliferation Index. Cytometry. Part a 95(7): 714-716, 2019

Quantum dots-based quantitative and in situ multiple imaging on ki67 and cytokeratin to improve ki67 assessment in breast cancer. Plos one 10(4): E0122734, 2015

Digital image analysis of Ki67 in hot spots is superior to both manual Ki67 and mitotic counts in breast cancer. Histopathology 72(6): 974-989, 2018

A Comparison of Visual Assessment and Automated Digital Image Analysis of Ki67 Labeling Index in Breast Cancer. Plos one 11(2): E0150505, 2016

Proliferation assessment in breast cancer: A double-staining technique for AgNOR quantification in MIB-1 positive cells especially adapted for image cytometry. Micron 31(2): 151-159, 2000

A methodology for comprehensive breast cancer Ki67 labeling index with intra-tumor heterogeneity appraisal based on hexagonal tiling of digital image analysis data. Virchows Archiv 2015:, 2015

Cytometric analysis of proliferation marker KI67 in epithelial cells of breast carcinoma identified with anti-cytokeratin antibody. Cytometry 59A(1): 138, 2004

A double-staining technique for AgNORs quantification in MIB1 positive cells adapted for image cytometry Application to proliferation assessment in breast cancer. Cytometry Suppl. (10): 150, 2000

Ki67 labelling index in gastric carcinomas. An immunohistochemical study using double staining for the evaluation of the proliferative activity of diffuse-type carcinomas. Journal of Pathology 182(1): 62-67, 1997

Practical approaches to automated digital image analysis of Ki-67 labeling index in 997 breast carcinomas and causes of discordance with visual assessment. Plos one 14(2): E0212309, 2019

Virtual Double Staining: A Digital Approach to Immunohistochemical Quantification of Estrogen Receptor Protein in Breast Carcinoma Specimens. Applied Immunohistochemistry and Molecular Morphology 26(9): 620-626, 2018

Comparison of cell proliferation in breast carcinoma using image analysis ki67 and flow cytometric systems. Laboratory Investigation 64(1): 123A, 1991

A methodology to ensure and improve accuracy of Ki67 labelling index estimation by automated digital image analysis in breast cancer tissue. Breast Cancer Research 16(2): R35, 2014

Cell proliferation in non-Hodgkin's lymphomas. Digital image analysis of Ki-67 antibody staining. American Journal of Pathology 134(2): 327-336, 1989