Hematoxylin-Eosin Staining Kit

Paraffin Section Staining

1. Deparaffinize sections in xylene I and xylene II for 5-10 min each.

2. Rehydrate in absolute ethanol I and II for 3 min each, followed by 95%, 85%, and 75% ethanol for 2 min each, and finally rinse with distilled water.

3. Stain the sections with Hematoxylin Staining Solution for 3-10 min.

4. Rinse under running tap water for 5-10 min to allow complete nuclear bluing. If necessary, briefly differentiate with a routine laboratory acid-alcohol solution and blue again.

5. Stain with Eosin Staining Solution for 1-3 min.

6. Rapidly dehydrate through 75%, 85%, and 95% ethanol, then absolute ethanol I and II for 2-3 min each, and clear in xylene I and II for 3-5 min each.

7. Mount with neutral mounting medium or another suitable mounting reagent and observe under a microscope.

Frozen Section Staining

1. Equilibrate frozen sections to room temperature, fix with 4% paraformaldehyde for 10-20 min, and wash 2-3 times with PBS.

2. Stain with Hematoxylin Staining Solution for 3-10 min and rinse under running tap water for 5-10 min.

3. Stain with Eosin Staining Solution for 1-3 min and briefly rinse with distilled water.

4. Dehydrate, clear, and mount as required.

Cultured Cell Staining

1. Remove the culture medium, wash the cells twice with PBS, and fix with 4% paraformaldehyde for 10-20 min.

2. Wash 2-3 times with PBS, add sufficient Hematoxylin Staining Solution to cover the cells, and stain for 3-10 min.

3. Rinse gently under running tap water until the background is clear and the nuclei are fully blued.

4. Add Eosin Staining Solution and stain for 30 s-3 min, then briefly rinse with distilled water.

5. Select an appropriate dehydration, clearing, and mounting method according to the slide or culture vessel used, and observe the staining.