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Digital Quantification of Epidermal Protein Expression in Paraffin-Embedded Tissue Using Immunohistochemistry
Current methods of assessing immunohistochemistry center on semiquantitative visual grading scales. More objective methods utilizing digital quantification offer superior precision and, presumably, higher confidence with image comparison. However, their cost often remains prohibitive, and there is little customizability to separate subsections of interest in the tissue. Here we describe a method using two open-source software programs to analyze the intensity and density of signals in immunohistochemistry-stained tissue sections that account for tissue heterogeneity and allow for direct comparison between two samples. This method allows for quantitative assessment of epidermal protein expression. We herein demonstrate this workflow using an epidermal stain to
thymic stromal lymphopoietin.
KeywordsColorimetric analysis Epidermis GIMP ImageJ Protein expression Quantitative immunohistochemistry
This work was supported in part by the International Pemphigus and Pemphigoid Foundation (Identifying novel pharmacologic targets in bullous pemphigoid: Unraveling the mechanisms of eosinophils) and a departmental grant from the University of California Irvine Department of Dermatology both to KTA.
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