Publication: Quantification of protein expression in cells and cellular
subcompartments on immunohistochemical sections
using a computer supported image analysis system
Authors
Braun, Martin ; Kirsten, Robert ; Rupp, Niels J. ; Moch, Holger ; Fend, Falko ; Wernert, Nicolas ; Kristiansen, Glen ; Perner, Sven
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Publisher
F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología
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DOI
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info:eu-repo/semantics/article
Description
Abstract
Quantification of protein expression based on
immunohistochemistry (IHC) is an important step for
translational research and clinical routine. Several
manual (‘eyeballing’) scoring systems are used in order
to semi-quantify protein expression based on
chromogenic intensities and distribution patterns.
However, manual scoring systems are time-consuming
and subject to significant intra- and interobserver
variability. The aim of our study was to explore, whether
new image analysis software proves to be sufficient as
an alternative tool to quantify protein expression. For
IHC experiments, one nucleus specific marker (i.e., ERG
antibody), one cytoplasmic specific marker (i.e.,
SLC45A3 antibody), and one marker expressed in both
compartments (i.e., TMPRSS2 antibody) were chosen.
Stainings were applied on TMAs, containing tumor
material of 630 prostate cancer patients. A pathologist
visually quantified all IHC stainings in a blinded
manner, applying a four-step scoring system. For digital
quantification, image analysis software (Tissue Studio
v.2.1, Definiens AG, Munich, Germany) was applied to
obtain a continuous spectrum of average staining
intensity. For each of the three antibodies we found a
strong correlation of the manual protein expression score
and the score of the image analysis software. Spearman’s
rank correlation coefficient was 0.94, 0.92, and 0.90 for
ERG, SLC45A3, and TMPRSS2, respectively (p<0.01).
Our data suggest that the image analysis software Tissue
Studio is a powerful tool for quantification of protein
expression in IHC stainings. Further, since the digital analysis is precise and reproducible, computer supported
protein quantification might help to overcome intra- and
interobserver variability and increase objectivity of IHC
based protein assessment.
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Citation
Histology and histopathology, Vol. 28, n.º 5 (2013)
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