Journal Article
. 2011 May; 2(4):187-94.
doi: 10.5306/wjco.v2.i4.187.

Automation of immunohistochemical evaluation in breast cancer using image analysis

Keerthana Prasad 1 Avani Tiwari  Sandhya Ilanthodi  Gopalakrishna Prabhu  Muktha Pai  
  • PMID: 21611095
  •     36 References
  •     3 citations


Aim: To automate breast cancer diagnosis and to study the inter-observer and intra-observer variations in the manual evaluations.

Methods: Breast tissue specimens from sixty cases were stained separately for estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor-2 (HER-2/neu). All cases were assessed by manual grading as well as image analysis. The manual grading was performed by an experienced expert pathologist. To study inter-observer and intra-observer variations, we obtained readings from another pathologist as the second observer from a different laboratory who has a little less experience than the first observer. We also took a second reading from the second observer to study intra-observer variations. Image analysis was carried out using in-house developed software (TissueQuant). A comparison of the results from image analysis and manual scoring of ER, PR and HER-2/neu was also carried out.

Results: The performance of the automated analysis in the case of ER, PR and HER-2/neu expressions was compared with the manual evaluations. The performance of the automated system was found to correlate well with the manual evaluations. The inter-observer variations were measured using Spearman correlation coefficient r and 95% confidence interval. In the case of ER expression, Spearman correlation r = 0.53, in the case of PR expression, r = 0.63, and in the case of HER-2/neu expression, r = 0.68. Similarly, intra-observer variations were also measured. In the case of ER, PR and HER-2/neu expressions, r = 0.46, 0.66 and 0.70, respectively.

Conclusion: The automation of breast cancer diagnosis from immunohistochemically stained specimens is very useful for providing objective and repeatable evaluations.

Keywords: Automation; Breast cancer diagnosis; Computer aided diagnosis; Image analysis; Immunohistochemical study.

Digital image analysis for the evaluation of the inflammatory infiltrate in psoriasis.
Amber Y Goedkoop, Menno A de Rie, +5 authors, Maarten C Kraan.
Arch Dermatol Res, 2005 Jul 14; 297(2). PMID: 16012877
Image analysis for the study of radionuclide distribution in tissue sections.
J L Humm, R M Macklis, +2 authors, L M Chin.
J Nucl Med, 1994 Jul 01; 35(7). PMID: 8014686
HER2 status in a population-derived breast cancer cohort: discordances during tumor progression.
Ulla Wilking, Eva Karlsson, +6 authors, Jonas Bergh.
Breast Cancer Res Treat, 2010 Jul 16; 125(2). PMID: 20628810
Prognostic ability of a panel of immunohistochemistry markers - retailoring of an 'old solution'.
Donal J Brennan, William M Gallagher.
Breast Cancer Res, 2008 Mar 12; 10(1). PMID: 18331621    Free PMC article.
Semi-automated imaging system to quantitate estrogen and progesterone receptor immunoreactivity in human breast cancer.
G M Sharangpani, A S Joshi, +5 authors, S H Barsky.
J Microsc, 2007 May 31; 226(Pt 3). PMID: 17535263
Image analysis of breast cancer immunohistochemistry-stained sections using ImageJ: an RGB-based model.
T Vrekoussis, V Chaniotis, +4 authors, O Zoras.
Anticancer Res, 2010 Jan 02; 29(12). PMID: 20044607
Correlation of quantitative digital image analysis with the glomerular filtration rate in chronic allograft nephropathy.
Montserrat M Diaz Encarnacion, Matthew D Griffin, +4 authors, Joseph P Grande.
Am J Transplant, 2004 Feb 21; 4(2). PMID: 14974947
Quantitative autoradiography of tyrosine hydroxylase immunoreactivity in the rat brain.
R Raisman-Vozari, E Hirsch, +5 authors, Y Agid.
J Neurochem, 1991 Oct 01; 57(4). PMID: 1680158
Quantitation of estrogen and progesterone receptors by immunocytochemical and image analyses.
D C Aziz.
Am J Clin Pathol, 1992 Jul 01; 98(1). PMID: 1615913
Automated quantitative analysis (AQUA) of in situ protein expression, antibody concentration, and prognosis.
Anthony McCabe, Marisa Dolled-Filhart, Robert L Camp, David L Rimm.
J Natl Cancer Inst, 2005 Dec 22; 97(24). PMID: 16368942
Application of photoshop-based image analysis to quantification of hormone receptor expression in breast cancer.
H A Lehr, D A Mankoff, +2 authors, A M Gown.
J Histochem Cytochem, 1997 Nov 14; 45(11). PMID: 9358857
Quantitation of immunocytochemical estrogen and progesterone receptor content in fine needle aspirates of breast carcinoma using the SAMBA 4000 image analysis system.
M Auger, R L Katz, +3 authors, H Fritsche.
Anal Quant Cytol Histol, 1993 Aug 01; 15(4). PMID: 8397648
Quantitative analysis of estrogen receptor heterogeneity in breast cancer.
Gina G Chung, Maciej P Zerkowski, +2 authors, David L Rimm.
Lab Invest, 2007 Mar 06; 87(7). PMID: 17334408
Immunohistochemical quantitation of androgen receptor expression using color video image analysis.
D Kim, C W Gregory, G J Smith, J L Mohler.
Cytometry, 1999 Nov 30; 35(1). PMID: 10554175
Automated selection of DAB-labeled tissue for immunohistochemical quantification.
Eric M Brey, Zahid Lalani, +4 authors, Charles W Patrick.
J Histochem Cytochem, 2003 Apr 22; 51(5). PMID: 12704205
A "quickscore" method for immunohistochemical semiquantitation: validation for oestrogen receptor in breast carcinomas.
S Detre, G Saclani Jotti, M Dowsett.
J Clin Pathol, 1995 Sep 01; 48(9). PMID: 7490328    Free PMC article.
Highly Cited.
Dilated cardiomyopathy is associated with significant changes in collagen type I/III ratio.
M Pauschinger, D Knopf, +5 authors, H P Schultheiss.
Circulation, 1999 Jun 03; 99(21). PMID: 10351968
Quantitative immunohistochemistry by measuring cumulative signal strength accurately measures receptor number.
Kristina A Matkowskyj, Randal Cox, Robert T Jensen, Richard V Benya.
J Histochem Cytochem, 2003 Jan 21; 51(2). PMID: 12533529
Application of immunohistochemistry to breast lesions.
I-Tien Yeh, Carolyn Mies.
Arch Pathol Lab Med, 2008 Mar 06; 132(3). PMID: 18318578
Semiautomated computer-assisted image analysis to quantify 3,3'-diaminobenzidine tetrahydrochloride-immunostained small tissues.
Sandra Leal, Carmen Diniz, +4 authors, Paula Fresco.
Anal Biochem, 2006 Aug 18; 357(1). PMID: 16914112
Semiquantitative study of the distribution of two cellular antigens by computer-directed color analysis.
J M Lamaziere, J Lavallee, C Zunino, J Larrue.
Lab Invest, 1993 Feb 01; 68(2). PMID: 7680084
Quantitative image analysis of immunohistochemical stains using a CMYK color model.
Nhu-An Pham, Andrew Morrison, +5 authors, David W Hedley.
Diagn Pathol, 2007 Mar 01; 2. PMID: 17326824    Free PMC article.
Immunodetection in fine-needle aspirates and multiparametric (SAMBA) image analysis. Receptors (monoclonal antiestrogen and antiprogesterone) and growth fraction (monoclonal Ki67) evaluation in breast carcinomas.
C Charpin, L Andrac, +5 authors, M Toga.
Cancer, 1989 Mar 01; 63(5). PMID: 2644015
Morphological detection and quantification of lipoprotein(a) deposition in atheromatous lesions of human aorta and coronary arteries.
A Niendorf, M Rath, +4 authors, M Dietel.
Virchows Arch A Pathol Anat Histopathol, 1990 Jan 01; 417(2). PMID: 2142355
18F-FDG PET of locally invasive breast cancer and association of estrogen receptor status with standardized uptake value: microarray and immunohistochemical analysis.
Joseph R Osborne, Elisa Port, +5 authors, Steven Larson.
J Nucl Med, 2010 Mar 20; 51(4). PMID: 20237034    Free PMC article.
Immunohistochemical evaluation of human epidermal growth factor receptor 2 and estrogen and progesterone receptors in breast carcinoma in Jordan.
Nidal M Almasri, Mohammad Al Hamad.
Breast Cancer Res, 2005 Sep 20; 7(5). PMID: 16168103    Free PMC article.
Objective measurement of breast cancer oestrogen receptor status through digital image analysis.
R Mofidi, R Walsh, +6 authors, N O'Higgins.
Eur J Surg Oncol, 2003 Feb 01; 29(1). PMID: 12559071
Estrogen receptor immunocytochemistry in paraffin embedded tissues with ER1D5 predicts breast cancer endocrine response more accurately than H222Sp gamma in frozen sections or cytosol-based ligand-binding assays.
L P Pertschuk, J G Feldman, +4 authors, C Axiotis.
Cancer, 1996 Jun 15; 77(12). PMID: 8640700
Theory and implementation of an electronic, automated measurement system for images obtained from immunohistochemically stained slides.
Gian Kayser, Dominik Radziszowski, +2 authors, Klaus Kayser.
Anal Quant Cytol Histol, 2006 Mar 29; 28(1). PMID: 16566277
Densitometric and morphometric study of immunocytochemical estrogen receptors detection in breast carcinomas.
G Santeusanio, A Mauriello, +4 authors, M Oberholzer.
Pathol Res Pract, 1992 Jun 01; 188(4-5). PMID: 1409075
Estrogen receptors in 699 primary breast cancers: a comparison of immunohistochemical and biochemical methods.
A Molino, R Micciolo, +7 authors, R Nortilli.
Breast Cancer Res Treat, 1995 Jun 01; 34(3). PMID: 7579486
Colour-texture based image analysis method for assessing the hormone receptors status in breast tissue sections.
Spiros Kostopoulos, Dionisis Cavouras, +6 authors, George Nikiforidis.
Annu Int Conf IEEE Eng Med Biol Soc, 2007 Nov 16; 2007. PMID: 18003125
Cytometrical image analysis for immunohistochemical hormone receptor status in breast carcinomas.
Yutaka Hatanaka, Kaoru Hashizume, +3 authors, Yoichi Tani.
Pathol Int, 2003 Oct 01; 53(10). PMID: 14516320
Comparison of quantification of histochemical staining by hue-saturation-intensity (HSI) transformation and color-deconvolution.
Arnout C Ruifrok, Ruth L Katz, Dennis A Johnston.
Appl Immunohistochem Mol Morphol, 2003 Mar 01; 11(1). PMID: 12610362
Immunohistochemical determination of estrogen and progesterone receptor content in human breast cancer. Computer-assisted image analysis (QIC score) vs. subjective grading (IRS).
W Remmele, K H Schicketanz.
Pathol Res Pract, 1993 Sep 01; 189(8). PMID: 8302707
Oestrogen receptor staining of paraffin-embedded breast carcinomas following short fixation in formalin: a comparison with cytosolic and frozen section receptor analyses.
W A Raymond, A S Leong.
J Pathol, 1990 Apr 01; 160(4). PMID: 2162939
Imaging as a diagnostic and therapeutic tool in clinical oncology.
Eddie Yin-Kwee Ng, Rajendra U Acharya.
World J Clin Oncol, 2011 May 26; 2(4). PMID: 21611092    Free PMC article.
IHC Profiler: an open source plugin for the quantitative evaluation and automated scoring of immunohistochemistry images of human tissue samples.
Frency Varghese, Amirali B Bukhari, Renu Malhotra, Abhijit De.
PLoS One, 2014 May 08; 9(5). PMID: 24802416    Free PMC article.
Highly Cited.
Quantitative Image Analysis of Human Epidermal Growth Factor Receptor 2 Immunohistochemistry for Breast Cancer: Guideline From the College of American Pathologists.
Marilyn M Bui, Michael W Riben, +11 authors, M Elizabeth Hammond.
Arch Pathol Lab Med, 2019 Jan 16; 143(10). PMID: 30645156    Free PMC article.