Overview
hematoxylin and eosin staining, abbreviated as HE staining, is the most widely used technique in histology, embryology and pathology. HE staining is the tissue sections or cell smears were immersed in hematoxylin staining solution and eosin staining solution, after appropriate time and processing, so that tissues, cells and other components stained with different shades of color, different refractive indices are generated to facilitate observation and study under an optical microscope. Hematoxylin is a kind of basic dye, which is extracted from hematoxylin and oxidized to form hematoxylin, which is used together with mordant, mainly in the nucleus of the staining Chemical Book color and cytoplasm of ribosomes with purple blue. The molecular formula of hematoxylin is C16H14O6. The commonly used hematoxylin dyeing solutions mainly include Ehrlich hematoxylin dyeing solution, Harris hematoxylin dyeing solution, Cole hematoxylin dyeing solution, Mayer hematoxylin dyeing solution, Carazzi hematoxylin dyeing solution, modified Lillie-Mayer hematoxylin dyeing solution and Gill modified hematoxylin dyeing solution. Eosin, an acid dye, is divided into two types: water-soluble and alcohol-soluble, which mainly makes the components in cytoplasm and extracellular matrix red.
after long-term use, it was found that the existing hematoxylin dyeing solution also has the following defects:
1) easy to produce metal film and dye precipitation, so as to contaminate the slice, and in the production process, the above stains on the slice are difficult to remove, resulting in uneven staining, even unable to read the film, need to re-production;
2) false Blue staining is easy to appear in the staining stage, and part of the cytoplasm is also stained blue, which affects the accuracy of the follow-up observation results.
preparation
hematoxylin dyeing solution, each of the hematoxylin dyeing solution includes the following components: hematoxylin 1g; Aluminum salt: aluminum ammonium sulfate 10g; Oxidant: Red mercury oxide 0.25g; Benzalkonium chloride 0.1g; Alcohol: ethanol 10ml; weak acid: glacial acetic acid 2.1g; Balance deionized water. Its preparation method is as follows: a. Taking alcohol and hematoxylin in a selected ratio, adding the hematoxylin to alcohol and dissolving to obtain a hematoxylin alcohol solution for use; B. Water and an aluminum salt are taken at a selected ratio so that the aluminum salt is completely dissolved in water, and the hematoxylin alcohol solution obtained in step a is added to obtain a mixed solution; c. The oxidant, benzalkonium chloride and weak acid are taken according to the selected proportion, the oxidant is stirred and added to the mixture obtained in Step B, the solution is heated to become dark purple, then cooled, and the benzalkonium chloride and weak acid are added, mixed evenly, filtered, that is, hematoxylin staining solution.