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Updated: Jun 4, 2026
Heterogeneity Mapping of Protein Expression in Tumors using Quantitative Immunofluorescence
Published on: October 25, 2011
Parametric mapping of immunohistochemically stained tissue sections; a method to quantify the colocalization of tumor
Saskia E Rademakers1, Paul F Rijken, Wenny J Peeters
1Department of Radiation Oncology, 874 Radboud University Nijmegen Medical Centre, P.O. Box 9101, Nijmegen, 6500 HB, The Netherlands. s.rademakers@rther.umcn.nl
Background:
Automated analysis of immunohistochemically stained tissue sections is of great importance in cancer research to detect tumor-specific prognostic markers and make therapy decisions. Here, an automated quantitative analysis is presented to assess the colocalization of CAIX, a membrane-bound hypoxic marker and Ki-67, a nuclear proliferation marker.
Methods:
Tissue sections of 104 biopsies from 89 patients were stained for CAIX and Ki-67 with diaminobenzidine and haematoxylin counterstain. Image scans of whole tumor sections were recorded and image maps were created with parametric mapping to quantify the markers and assess the colocalization.
Results:
The fraction of CAIX showed a range of 0-93%. The interobserver correlation and the correlation between manual scores and automated analysis were both very strong (rs=0.96, p<0.0001, and rs=0.97, p<0.0001). The labelling index of Ki-67 exhibited a range of 0-42% with less strong interobserver and manual to automated analysis correlations (rs=0.90, p<0.0001, and rs=0.71, p<0.0008). The relative tumor area positive for both markers varied from 0-76%.
Conclusion:
Parametric mapping of immunohistochemically stained tumor sections is a reliable method to quantitatively analyze membrane-bound proteins and assess the colocalization of various tumor markers in different subcellular compartments.
