ImmunoRatio: a publicly available web application for quantitative image analysis of estrogen receptor (ER), progesterone receptor (PR), and Ki-67

Tuominen, Vilppu J.; Ruotoistenmäki, Sanna; Viitanen, Arttu; Jumppanen, Mervi; Isola, Jorma

doi:10.1186/bcr2615

Cited by 424 publications

(391 citation statements)

References 21 publications

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“…Cells were washed with PBS prior to blocking endogenous peroxidases with 0.3% (v/v) Images were observed using a Zeiss Axioskop light microscope, AxioCam ICc 3 Zeiss camera and AxioVision System Software (release 4.6; Carl Zeiss Imaging Solutions, Munchen, Germany). Image analysis was performed using the ImmunoRatio automated image analysis program (Tuominen et al 2010) as a plugin in Fiji (ImageJ; http://fiji.sc/How_to_ cite_Fiji%3F) (Schindelin et al 2012). Three fields of view per sorted cell sub-population were analysed.…”

Section: Immunocytochemistry and Image Analysismentioning

confidence: 99%

Common fibroid-associated genes are differentially expressed in phenotypically dissimilar cell populations isolated from within human fibroids and myometrium

Holdsworth-Carson¹,

Zaitseva²,

Girling³

et al. 2014

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Uterine fibroids are a prevalent gynaecological condition in reproductive-aged women and are the commonest reason for hysterectomy. The cellular composition of clonal fibroids are heterogeneous, with phenotypically dissimilar cells that include smooth muscle cells (SMC), vascular SMC (VSMC) and fibroblasts. The aim of our study was to investigate genes that are commonly differentially expressed between fibroid and myometrial whole tissues in phenotypically different sub-populations of cells isolated from fibroid and myometrium. Genes to be investigated by fluorescence-activated cell sorting, quantitative real-time PCR and immunocytochemistry include transforming growth factor b (TGFB) and retinoic acid (RA) signalling families and steroid hormone receptors. We hypothesised that each cell population isolated from fibroid and myometrium would differ in the expression of fibroid-associated genes. We demonstrated that phenotypically different cellular constituents of uterine fibroids differentially express cellular RA-binding protein 2 (CRABP2), progesterone receptor B (PRB) and TGFB receptor 2 mRNA in fibroid-derived cells of VSMC and SMC phenotype. CRABP2 mRNA was also differentially expressed in fibroblasts and VSMC sub-populations from within clonal fibroid tumours. We conclude that differential regulation of RA, TGFB and PR pathway transcription occurs in fibroid-associated SMC and -fibroblasts and that investigation of paracrine interactions between different cell types within the fibroid microenvironment provides an important new paradigm for understanding the pathophysiology of this common disease.

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Section: Immunocytochemistry and Image Analysismentioning

confidence: 99%

Common fibroid-associated genes are differentially expressed in phenotypically dissimilar cell populations isolated from within human fibroids and myometrium

Holdsworth-Carson¹,

Zaitseva²,

Girling³

et al. 2014

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“…have also been reported 18 . One study found pathologist vs. pathologist kappa agreements of 0.57 for ER and 0.51 for PR, however this value dropped to 0.42 for ER and 0.36 for PR when comparing pathologists from different laboratories 19 .…”

Section: Discussionmentioning

confidence: 53%

Simultaneous automatic scoring and co‐registration of hormone receptors in tumor areas in whole slide images of breast cancer tissue slides

et al. 2016

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Automation of downstream analysis may offer many potential benefits to routine histopathology. One area of interest for automation is in the scoring of multiple immunohistochemical markers to predict the patient's response to targeted therapies. Automated serial slide analysis of this kind requires robust registration to identify common tissue regions across sections. We present an automated method for co‐localized scoring of Estrogen Receptor and Progesterone Receptor (ER/PR) in breast cancer core biopsies using whole slide images. Regions of tumor in a series of fifty consecutive breast core biopsies were identified by annotation on H&E whole slide images. Sequentially cut immunohistochemical stained sections were scored manually, before being digitally scanned and then exported into JPEG 2000 format. A two‐stage registration process was performed to identify the annotated regions of interest in the immunohistochemistry sections, which were then scored using the Allred system. Overall correlation between manual and automated scoring for ER and PR was 0.944 and 0.883, respectively, with 90% of ER and 80% of PR scores within in one point or less of agreement. This proof of principle study indicates slide registration can be used as a basis for automation of the downstream analysis for clinically relevant biomarkers in the majority of cases. The approach is likely to be improved by implantation of safeguarding analysis steps post registration. © 2016 International Society for Advancement of Cytometry

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“…15 We found that for all three techniques of MIB-1 quantification, there was no statistically significant difference in MIB-1 indices between biopsies and resection specimen: 20% vs 22% for visually estimated data (Figure 2, left panels), 22% vs 22% for counted data, and 14% vs 14% for image analysis (IMMUNORATIO), respectively. At the same time, the ratio of MIB-1 over mitotic figures was significantly (Po0.02) reduced in resection specimen (Figure 2, right panel).…”

Section: Mitoses Overestimated In Resectionsmentioning

confidence: 71%

“…Using inbuild map commands, this software identifies the surface covered by hematoxylin-counterstained (blue) nuclei and the surface covered by immunoreactive (brown) nuclei, and calculates an index of immunostained cells over the total cell number. 15 …”

Section: Immunohistochemistrymentioning

confidence: 99%

Mitotic figure counts are significantly overestimated in resection specimens of invasive breast carcinomas

et al. 2013

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Several authors have demonstrated an increased number of mitotic figures in breast cancer resection specimen when compared with biopsy material. This has been ascribed to a sampling artifact where biopsies are (i) either too small to allow formal mitotic figure counting or (ii) not necessarily taken form the proliferating tumor periphery. Herein, we propose a different explanation for this phenomenon. Biopsy and resection material of 52 invasive ductal carcinomas was studied. We counted mitotic figures in 10 representative high power fields and quantified MIB-1 immunohistochemistry by visual estimation, counting and image analysis. We found that mitotic figures were elevated by more than three-fold on average in resection specimen over biopsy material from the same tumors (20 ± 6 vs 6 ± 2 mitoses per 10 high power fields, P ¼ 0.008), and that this resulted in a relative diminution of post-metaphase figures (anaphase/telophase), which made up 7% of all mitotic figures in biopsies but only 3% in resection specimen (Po0.005). At the same time, the percentages of MIB-1 immunostained tumor cells among total tumor cells were comparable in biopsy and resection material, irrespective of the mode of MIB-1 quantification. Finally, we found no association between the size of the biopsy material and the relative increase of mitotic figures in resection specimen. We propose that the increase in mitotic figures in resection specimen and the significant shift towards metaphase figures is not due to a sampling artifact, but reflects ongoing cell cycle activity in the resected tumor tissue due to fixation delay. The dwindling energy supply will eventually arrest tumor cells in metaphase, where they are readily identified by the diagnostic pathologist. Taken together, we suggest that the rapidly fixed biopsy material better represents true tumor biology and should be privileged as predictive marker of putative response to cytotoxic chemotherapy.

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ImmunoRatio: a publicly available web application for quantitative image analysis of estrogen receptor (ER), progesterone receptor (PR), and Ki-67

Cited by 424 publications

References 21 publications

Common fibroid-associated genes are differentially expressed in phenotypically dissimilar cell populations isolated from within human fibroids and myometrium

Common fibroid-associated genes are differentially expressed in phenotypically dissimilar cell populations isolated from within human fibroids and myometrium

Simultaneous automatic scoring and co‐registration of hormone receptors in tumor areas in whole slide images of breast cancer tissue slides

Mitotic figure counts are significantly overestimated in resection specimens of invasive breast carcinomas

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