"How do we accurately calculate and normalize IHC staining percentages when different tissue components (epithelial&c.t) occupy unequal area?

01 January 1970 0 3K Report

"When analyzing immunohistochemistry (IHC) stained images, calculating the percentage of positive staining in epithelial or connective tissue based solely on the area occupied in the field of view may be misleading. This is because if one component (e.g., connective tissue) naturally occupies less space in the field compared to another component (e.g., epithelial tissue), its final percentage calculation will be disproportionately low.

Questions:

How should we standardize or normalize these measurements to account for the different baseline amounts of tissue components?

What would be an appropriate ratio calculation method to ensure fair comparison between tissue components that occupy different proportions of the field of view?

What is the best practice for quantifying staining when tissue components have unequal spatial distribution?"

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