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Updated: Apr 10, 2026

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
Published on: April 8, 2016
Strategies to improve quantitative assessment of immunohistochemical and immunofluorescent labelling
Sarah J Johnson1, F Rohan Walker2
11] School of Electrical Engineering and Computer Science, University of Newcastle, Callaghan, NSW, Australia [2] Hunter Medical Research Institute, New Lambton Heights, NSW, Australia.
Abstract:
Binary image thresholding is the most commonly used technique to quantitatively examine changes in immunolabelled material. In this article we demonstrate that if implicit assumptions predicating this technique are not met then the resulting analysis and data interpretation can be incorrect. We then propose a transparent approach to image quantification that is straightforward to execute using currently available software and therefore can be readily and cost-effectively implemented.
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