Hematoxylin is a deep blue-purple color basic dye which has high affinity to acidic parts of cells, such as the nucleus.

Store at 4°C.

Mayer’s Hematoxylin Preparation:

a. Add 50 g aluminum potassium sulfate in 1L ddH₂O. Mix well until all aluminum potassium sulfate dissolved.

b. Add 1 g hematoxylin into the aluminum potassium sulfate solution and completely dissolve it.

c. Add 0.2 g sodium iodate and 20 ml acetic acid into the solution.

d. Bring the solution to boil and cool.

e. Filter the soluition .

f. Store at 4°C.

Note: For Mayer’s haematoxylin contains lower concentration of dyes which can selectively stain cell nucleus, you may get a light staining and lower background. If you want deep blue staining, you may choose Harris Hematoxylin.

Hematoxylin staining:

1. Immerse the slides (slides should be fixed in either alcohol or an aldehyde-based fixative) for 30 sec with agitation by hand in H₂O. This step is to hydrate the tissue and prevent hematoxylin to precipitate with salts and buffers.

2. Immerse the slides into Mayer’s hematoxylin and agitate for 8 min.

3. Wash in warm running tap water

Hematoxylin is a deep blue-purple color basic dye which has high affinity to acidic parts of cells, such as the nucleus. Hematoxylin is used to illustrate nucleus in tissue staining, like H&E staining. There are three different kinds of Hematoxylin, Ehrlich’s haematoxylin, Harris’s haematoxylin and Mayer’s haematoxylin. Mayer’s haematoxylin contains lower concentration of dyes which can selectively stain cell nucleus. Thus, it can be used as a counterstain for many immunohistochemical or hybridization procedures.

[1] Fischer AH, Jacobson KA, Rose J, Zeller R. Hematoxylin and eosin staining of tissue and cell sections. CSH Protoc. 2008 May 1;2008:pdb.prot4986. doi: 10.1101/pdb.prot4986. PMID: 21356829.