The Herovici staining method was proposed in 1963 and can be used to differentiate between young and mature collagen proteins. The principle is similar to Masson's trichrome method, using a combination of multi-color dyes for dyeing. Depending on the density of different types of collagen, the affinity with dyes of different sizes and molecular weights also varies. Red dyes with smaller molecular weights are more likely to bind to mature (dense) collagen, dyeing it pink to purple red. Blue dyes with larger molecular weights are more likely to bind to young (loose) collagen, dyeing it blue to blue purple.

The Herovici Collagen Stain Kit consists of Weigert Iron Hematoxylin Working Solution and Herovici Staining Solution, which can stain cell nucleus blue to black. Herovici Staining Solution contains a yellow staining agent, which can stain red blood cells and muscle fibers yellow.

 

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