Preparation before experiment

Bring your own hematoxylin differentiation solution (recommended G1039), hematoxylin re-blue solution (recommended G1040), xylene, gradient ethanol, absolute ethanol, neutral gum, etc.



Instructions

1. Paraffin sections are deparaffinized to water: sections are sequentially deparaffinized with xylene for 10 minutes, replaced with fresh xylene for 10 minutes, absolute ethanol for 5 minutes, fresh absolute ethanol for 5 minutes, 90% ethanol for 5 minutes, and 75% ethanol 5 min, wash with tap water.

2. The section is immersed in PAS staining solution B for 10-15 min, and then immersed in pure water 3 times, each time is about 10 s;

3. The sections are placed in PAS staining solution A (return to room temperature in advance), covered and protected from light for 25-30 min, and rinsed with running water for 5 min.

4. (Optional) The section is stained in PAS staining solution C for 30 s, and washed with tap water. Then the slices were differentiated with hematoxylin differentiation solution for 3 s, and then washed with tap water; after hematoxylin-returning solution was returned to blue for 3 s, washed with tap water. This step is to stain the nucleus.

5. Cut slices into three cylinders of absolute ethanol for dehydration, each time for 5 minutes; pass xylene to clear for 5 minutes, replace with fresh xylene and then clear for 5 minutes, then cover with neutral gum.


Precautions

1. PAS staining solution B can be reused. The staining solution is transparent. If it is obviously yellow, there are impurities, or the staining of the tissue is too light, it needs to be replaced with a new one.

2. PAS staining solution A needs to be stored at 4°C, take it out and return to room temperature before use. When the color of the solution is obviously red, it is recommended to replace with a new dye solution.

3. Please wear lab coat and disposable gloves during operation.