Hematoxylin and Eosin (H&E) Staining Kit (with Differentiation and Bluing Reagents) 

Hematoxylin and Eosin (H&E) Staining Kit (with Differentiation and Bluing Reagents) can be used for staining paraffin sections, frozen sections, and tissue cells. After hematoxylin-eosin combined staining, the cytoplasm appears red or pink, and the cell nucleus appears blue or bluish-purple.

Reagent Preparation
1. Fixative: 4% paraformaldehyde fixative
2. Differentiation solution: 5% acetic acid solution or 0.5% hydrochloric acid ethanol
3. Other reagents: xylene, mounting medium, anhydrous ethanol, 90% ethanol, 80% ethanol, 70% ethanol, distilled water
Sample Preparation
1. Paraffin Section Preparation
1) Dewaxing in xylene for 10 min;
2) Dewaxing with fresh xylene for 10 min;
3) Dewaxing with xylene:anhydrous ethanol = 1:1 for 5 min;
4) Dewaxing with 90% ethanol for 2 min;
5) Dewaxing with 80% ethanol for 2 min;
6) Dewaxing with 70% ethanol for 2 min;
7) Dewaxing with distilled water for 2 min.

2. Frozen Section Treatment
1) Fix in fixative for at least 10 min;
2) Wash with distilled water for 2 min.

3. Cultured Cell Treatment
1) Fix in fixative for at least 10 min;
2) Wash with distilled water for 2 min;
3) Repeat washing with distilled water once.

Observation
1. Direct Observation
Observe directly after staining.
2. Observation after Mounting
1) Wash with 95% ethanol for 3 min, then wash with anhydrous ethanol for 3 min;
2) Clear with xylene for 10 min;
3) Mount with neutral resin or mounting medium and observe.

Note:
1. Differentiation after staining is an optional step. Differentiation results in clearer nuclear staining, but the time should not be too long.
2. The staining solution can be reused; replace it with a new solution only when the staining performance significantly decreases.
3. Ethanol is volatile; it is recommended to prepare and use the series of ethanol solutions immediately.
4. Hematoxylin staining is difficult at lower temperatures; the staining time can be appropriately extended.
5. The dewaxing time can be appropriately shortened at higher room temperatures.
6. It is not recommended to perform other fluorescent staining, such as immunofluorescence, after staining with this kit.
7. This product is for scientific research use by professionals only and must not be used for clinical diagnosis or treatment, nor for food or pharmaceuticals.
8. For your safety and health, please wear a lab coat and disposable gloves when handling this product.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

[Hematoxylin and Eosin (H&E) Staining Kit (with Differentiation and Bluing Reagents)]

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.