Throughout this week in the Histology lab, I was operating the automated hematoxylin and eosin stainer, while also submitting stained slides to the pathologists to be reviewed.
Hematoxylin and eosin is a stain used in routine Histology slides. The hematoxylin stains nuclei in the tissue blue, whereas eosin stains the cytoplasm and other structures varying shades of pink. Hematoxylin is a natural dye originating from the logwood tree, Haematoxylum campechianum. However, it is not hematoxylin that is truly staining the nuclei, but rather its oxidized state of hematein. Oxidation of hematoxylin to hematein can occur using two different methods. One is a natural oxidation process that uses air or light. This process takes a significant amount of time to oxidize the dye, however, it produces a very stable stain. The other method uses chemical oxidation that requires either sodium iodate or potassium iodate. While this process occurs much quicker than the natural method, these stains may deteriorate and require filtering before use. However, this oxidized state of hematoxylin is still not sufficient to stain tissues. Hematoxylin must also be combined with a ‘mordant’, which gives the dye affinity to the tissue. Typical mordants that are added to hematoxylin are aluminum, iron, lead, tungsten, and molybdenum. Continue reading