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The page below is a sample from the LabCE course Immunohistochemistry (IHC) Basics in Histology. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Chromogen Enhancers

Chromogen enhancers increase the intensity of the color appearance of positive IHC staining reactions. 3,3’-diaminobenzidine (DAB) enhancers contain compounds that continue the DAB reaction or darken/change the color. An enhancer to the color can be heavy metals, such as copper, silver, nickel, gold, or cobalt. The most commonly used metal as an enhancer is nickel, which gives a blue-black color to the DAB. For electron microscopy IHC procedures, gold is used.
In any case, when using an enhancer for the first time, a control slide using the DAB without enhancers should be stained prior to adding the enhancer. The presence of the original unenhanced reaction validates that the enhancer is not turning a negative stain result into a positive stain result.