The acidic nuclei in cells shows high affinity with basic dye (haematoxylin), while the cytoplasm containing alkali substances which show high affinity with acid dye (eosin). When stained with Haematoxylin-Eosin Solution, the nuclei of cells or tissue section shows sharp blue, and the cytoplasm, collagen and muscle fibers show the color of pink to red in different extent.
1.Deparaffinize sections, 2 changes of xylene, 20 minutes each. Dehydrate in 2 changes of absolute alcohol, 5 minutes each. Rinse 75% alcohol for 5 minutes. Wash briefly in tap water.
2.Stain in hematoxylin solution for 3-5 minutes. Differentiate in acid alcohol Then bluing in ammonia water, wash in running tap water for 5 minutes.
3.Dehydrate through 85% alcohol and 95% alcohol respectively. Counterstain in Eosin Y solution (alcohol) for 5 minutes.
4.Dehydrate in 3 changes of absolute alcohol, 5 minutes each. Clear in 2 changes of xylene, 5 minutes each. Mount with resin mounting medium.
5.Microscopy examination and analysis
Nuclei should be blue, cytoplasm pink red. The nucleoplasm is clear and contrasting.
1. Low temperature is not good for hematoxylin coloring. You should extend the staining time under low indoor temperature.
2. The time for deparaffinize should be adjust according to room temperature. If room temperature is higher than 30℃, shorten the deparaffinize time.
3. The aim of differentiate is to increase the contrast. So the differentiate time should be strictly controlled.
4. For the smear sample or printed slice, start the H&E staining from step 5.
5. Please use the product in the shelf life. In order to better preserve the product, heat, low temperature and light should be avoid.
6. For your safety and health, please wear the lab coat and wear a disposable glove operation.