This method of staining is based on the fact that tissue structures bind differently to different dyes. The dye hematoxylin stains basophilic structures blue-violet, while eosin stains acidophilic structures pink. Basophilic structures usually include parts containing nucleic acids, such as ribosomes, the nucleus, and ribonucleic acid (RNA)-rich regions of the cytoplasm.
Operation method
basic program
Materials and Instruments
Slides Move 1. Place the developed slide (in a slide holder) into a staining tray with water. Caveat 1. If immersed in 0.1% ammonium hydroxide for too long, the radioactive autoradiographic latex will detach from the slide. 2. If the staining is too dark, the slide can be immersed in 50% ethanol and decolorized for a few moments (observe the dye diffusion condition), if the staining is too weak, it can be re-stained. For more product details, please visit Aladdin Scientific website.
Hematoxylin dye Ammonium hydroxide Eosin dye
Water bath Incubator
2. Prepare each of the following groups of liquids in the staining tray by number.
(1) Hematoxylin Staining Solution
(2) Water
(3) Water
(4) 0.1% NH4OH
(5) Water
(6) Water
(7) Eosin stain
(8) 95% ethanol
(9) 100% ethanol
(10) 100% ethanol
(11) Xylene
(12) Xylene
(13) Xylene3. Stain slides in hematoxylin staining solution for 20-30 s (do not stain too deeply), and immerse in water twice for 2 min each time.
4. Quickly immerse in 0.1% ammonium hydroxide and then in water.
5. Wash with water for 5 min, then stain with eosin for 20-30 s. '6. Dip the slide 8 times in the same dish of 95% ethanol, then 8 times in 100% ethanol, then 2 min in 100% ethanol for complete decolorization, and equilibrate in xylene 3 times for 2 min each.
Figure 1: Schematic diagram of slide fixation with fixation cutoff
7. Operate according to basic program 1, steps 5 to 9, for solid sheet processing.