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The most critical step in the Gomori-Wheatley trichrome procedure is the differentiation phase, where the tissue is exposed to a differentiating solution, typically containing hydrochloric acid and ethanol. This step is crucial because it selectively removes excess staining and allows for the clear visualization of collagen fibers and other tissue components. Proper differentiation ensures that the staining highlights the desired structures without over-staining or under-staining, which can compromise the analysis of the tissue.

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