Van Gieson Stain

Van Gieson Stain

Principle of Van Gieson Stain

The Van Gieson stain method involves two main components: acid fuchsin and picric acid. Acid fuchsin, with its medium to large molecular weight, selectively penetrates collagen fibers characterized by looser tissue structures, resulting in a red to pink stain. Picric acid, on the other hand, has a small molecular weight that enables it to permeate muscle fibers, glial cells, and red blood cells. These structures, with their dense tissue composition, are stained yellow by the picric acid.

Components and Reagents

The following are the primary reagents required for the Van Gieson staining process:

  • Weigert’s Iron Hematoxylin Solution A: Contains hematoxylin, a dye used to stain nuclei.
  • Weigert’s Iron Hematoxylin Solution B: Contains ferric chloride, an essential component for hematoxylin staining.
  • Van Gieson Stain: Comprising ferric chloride and other constituents necessary for the dual staining process.

Sample Preparation and Procedure

  • Ensure that the tissue section is thoroughly fixed.
  • Dewax the tissue slices and wash them in distilled water.
  • Apply Weigert’s Iron Hematoxylin Solution A followed by a mixture of Weigert’s Iron Hematoxylin Solution B (in a 1:1 ratio). This step stains nuclei and lasts for 5 to 10 minutes.
  • Rapidly differentiate the sample using 1% hydrochloric acid alcohol and wash it.
  • Stain the sample with Van Gieson stain for 1 to 2 minutes.
  • Immediately differentiate the sample using 95% alcohol.
  • Proceed with conventional dehydration, transparency, and seal using neutral resin.

Result Interpretation

  • Collagen fibers: Stained in red.
  • Muscle fibers, Nerve glial, Erythrocytes: Stained in yellow.
  • Nuclei: Stained in purple-black.


  • Mix Weigert’s Iron Hematoxylin Solution A and B prior to use to avoid oxidation and loss of staining power.
  • Differentiate the tissue swiftly in 95% ethanol after Van Gieson stain to prevent excessive discoloration.
  • Since acid fuchsin is soluble in water, do not rinse the sample in water after VG staining. Instead, differentiate using 95% ethanol followed by dehydration in absolute ethanol.
  • Professional expertise is recommended for using this reagent and interpreting results. Adhere to the instruction manual, use within the validity period, and practice personal hygiene protection.
  • Dispose of waste according to hospital or environmental protection guidelines.
  • Production date, batch number, and expiration date can be found on the outer packaging.


The Van Gieson stain technique offers a valuable means of visualizing and differentiating tissues under a microscope. By utilizing the unique staining properties of acid fuchsin and picric acid, this method highlights collagen and muscle fibers, providing insights into tissue composition and structure. Proper execution of the staining procedure, in accordance with the specified precautions, ensures accurate and informative results for histological analysis.

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