Quantitative image analysis of immunohistochemical stains using a CMYK color model

Pham, Nhu-An; Morrison, Andrew; Schwock, Joerg; Aviel‐Ronen, Sarit; Iakovlev, Vladimir V.; Tsao, Ming‐Sound; Ho, J. C.; Hedley, David W.

doi:10.1186/1746-1596-2-8

Cited by 153 publications

(58 citation statements)

References 24 publications

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“…pSTAT3 expression was scored as described previously: [13]. In brief, immunostaining scores (IHC score: 0-300) was calculated as the products of the staining intensity (1 (weak), 2 (moderate) or 3 (strong) expression) and the staining rates (percentages of tumor cells showing positive staining, 0-100 %).…”

Section: Methodsmentioning

confidence: 99%

Phosphorylation of signal transducer and activator of transcription 3 (STAT3) correlates with Her-2 status, carbonic anhydrase 9 expression and prognosis in esophageal cancer

Schoppmann

Jesch

Friedrich

et al. 2012

Clin Exp Metastasis

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The signal-transcriptional factor signal transducer and activator of transcription (STAT3) is overexpressed in various tumor entities and promotes tumor progression and metastasis. The tyrosine-kinase receptor Her-2 was shown to activate STAT3-expression and is overexpressed in a subtype of esophageal cancer (EC). STAT3 also regulates carbonic anhydrase IX (CAIX) expression in vivo, promoting IL-6-dependent tumor invasion.Tumor-tissue from 324 patients with EC and 45 patients with precursor lesions were analyzed for the expressions of tyrosine-705 phosphorylated STAT3 (pSTAT3). Data on Her-2-status and CAIX expression were available from previous studies. pSTAT3 was overexpressed in 40 % of adenocarcinomas (AC) and 50 % of squamous cell carcinomas of the esophagus and was associated with worse overall (OS) (p = 0.001) and disease free survival (DFS) (p = 0.001). In Barrett's mucosa without/low-grade dysplasia, pSTAT3 was expressed in 14 % compared to 27 % in patients with high-grade dysplasia (p = 0.018). A significant association between Her-2 and pSTAT3 was found in ACs (p = 0.013), showing that patients with tumors expressing both proteins have significantly worse OS (p = 0.0039) and DFS (p = 0.029). One hundred-fifty (46.3 %) cancer cases were considered as positive for CAIX expression, and a strong correlation between pSTAT3 and CAIX expression was seen (p < 0.001). pSTAT3 plays an important role in the development of AC. Expression of pSTAT3 correlates with Her-2 status and CAIX expression and is associated with tumor progression and worse outcome, offering expectantly therapeutical implications.

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Section: Methodsmentioning

confidence: 99%

Phosphorylation of signal transducer and activator of transcription 3 (STAT3) correlates with Her-2 status, carbonic anhydrase 9 expression and prognosis in esophageal cancer

Schoppmann

Jesch

Friedrich

et al. 2012

Clin Exp Metastasis

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“…When the results are negative in both original and diluted feces it might show that no HGF with affinity to DS is present in feces. The CMYK system was used for interpretation of colors as follows: 701500, 502500, 453000 are ranges of positive result and 354000, 254000 as negative results [15].…”

Section: Assay Accomplishment and Interpretation Of Resultsmentioning

confidence: 99%

A methachromatic-based experimental model for identification of bowel as the focus of an acute inflammation

Nakka¹,

Johansson²,

Shahzad³

et al. 2013

OJGas

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Diarrhea is the most common symptom of acute inflammation in gastrointestinal tract and the patients are isolated in order to inhibit transmission and to conduct investigations. Yet there is no standard test to distinguish gastrointestinal infection from more generalized diseases at admittance which might cause delay in therapy. Hepatocyte growth factor (HGF) is produced upon injury by mesenchymal cells. On the contrary to chronic inflammation, HGF produced in the course of acute inflammation is biologically active and shows binding affinity to heparan sulphate proteoglycan (HSPG) and dextran sulphate (DS). Based on this phenomenon, an agarose gel containing DS was prepared and immobilized on loops to investigate the feces samples for the presence or absence of growth factors such as HGF with affinity to DS. The study is conducted as a clinical evaluation of an experimental model to distinguish acute infectious gastroenteritis from other causes of diarrhea. 656 fecal samples gathered consequently from patients seeking for bowel disturbances and healthy were tested by the test and the medical reports were investigated. Upon interaction with DS, methylene blue changes color to pink. This phenomenon was inhibited by HGF and converted by addition of anti-HGF antibodies to the samples. The test distinguished acute infectious gastroenteritis with high sensitivity and specificity (96% and 92% respectively) from other causes of diarrhea. We introduce a metachromatic experimental model that might distinguish acute inflammation in alimentary tract from other causes of diarrhea. This model might be used in developing rapid diagnostic tests.

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“…Quantitation of immunohistochemistry was performed using ImageJ software (NIH). The yellow channel was extracted from the CMYK color scheme in order to obtain the best contrast and intensity scale of the image, and to eliminate background as much as possible [24]. The immunohistochemistry was quantified in four regions of the basilar bifurcation: the periapical region with high WSS and high, positive WSSG, the left posterior cerebral artery, the right posterior cerebral artery and the basilar artery.…”

Section: Methodsmentioning

confidence: 99%

Cellular and Molecular Responses of the Basilar Terminus to Hemodynamics during Intracranial Aneurysm Initiation in a Rabbit Model

et al. 2011

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Background/Aims: Hemodynamics constitute a critical factor in the formation of intracranial aneurysms. However, little is known about how intracranial arteries respond to hemodynamic insult and how that response contributes to aneurysm formation. We examined early cellular responses at rabbit basilar termini exposed to hemodynamic insult that initiates aneurysmal remodeling. Methods: Flow in the basilar artery was increased by bilateral carotid artery ligation. After 2 and 5 days, basilar terminus tissue was examined by immunohistochemistry and quantitative PCR. Results: Within 2 days of flow increase, internal elastic lamina (IEL) was lost in the periapical region of the bifurcation, which experienced high wall shear stress and positive wall shear stress gradient. Overlying endothelium was still largely present in this region. IEL loss was associated with localized apoptosis and elevated expression of matrix metalloproteinases (MMPs) 2 and 9. A small number of inflammatory cells were sporadically scattered in the bifurcation adventitia and were not concentrated in regions of IEL loss and MMP elevation. Elevated MMP expression colocalized with smooth muscle α-actin in the media. Conclusion: The initial vascular response to aneurysm-initiating hemodynamic insult includes localized matrix degradation and cell apoptosis. Such destructive remodeling arises from intrinsic mural cells, rather than through inflammatory cell infiltration.

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Quantitative image analysis of immunohistochemical stains using a CMYK color model

Abstract: Background: Computer image analysis techniques have decreased effects of observer biases, and increased the sensitivity and the throughput of immunohistochemistry (IHC) as a tissue-based procedure for the evaluation of diseases.

Cited by 153 publications

References 24 publications

Phosphorylation of signal transducer and activator of transcription 3 (STAT3) correlates with Her-2 status, carbonic anhydrase 9 expression and prognosis in esophageal cancer

Phosphorylation of signal transducer and activator of transcription 3 (STAT3) correlates with Her-2 status, carbonic anhydrase 9 expression and prognosis in esophageal cancer

A methachromatic-based experimental model for identification of bowel as the focus of an acute inflammation

Cellular and Molecular Responses of the Basilar Terminus to Hemodynamics during Intracranial Aneurysm Initiation in a Rabbit Model

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