Hematoxylin-eosin staining, called HE staining, one of the commonly used staining methods in paraffin section technology. 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 HE Solution, the nuclei of cells or tissue section shows sharp blue, and the cytoplasm, collagen and muscle fibers show the color pink to red in different extent. HE staining is the most basic and widely used technique method in histology, embryology, pathology and research.
1.Deparaffinize and rehydrate: sections were placed in xylene I 20minutes-xylene II 20 minutes-Absolute ethanol I 5 min-Absolute ethanol II 5 min-75% alcohol for 5 min, then wash in tap water.
2.Hematoxylin staining: stain in hematoxylin solution for 3-5min, tap water washing, Differentiate in acid alcohol, tap water washing, return blue liquid back to blue, running water washing.
3. Eosin staining:Dehydrate through 85% alcohol and 95% alcohol respectively for 5 min , and counterstain in Eosin Y solution (alcohol) for 5 min.
4.Dehydrated and Mount: dehydrate in absolute alcohol for 3 times ,5min each time ,clear in xylene for 2 times , 5min each time , mount with neutral gum.
5.Microscopy detection, image acquisition and analysis.
Nucleus should be blue, cytoplasm pink red. The cytoplasmis 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.