Hematoxylin-eosin (HE) staining
Dye preparation:
1. Hematite dyeing solution
(1) Weigh 0.5g hematoxylin, 5.0g ammonium or potassium hydrazine and 0.1g sodium iodate and dissolve in 70ml distilled water;
(2) Add 30 ml of glycerin and 2 ml of glacial acetic acid, mix well and filter to form a mother liquor;
(3) The mother liquor can be stored for a long time. The mother liquor is diluted 1:20 with distilled water to form a working fluid. The working fluid is also stored for a long time, but it should be filtered before each dyeing to remove the oxide film;
2. Yihong dyeing solution
Eosin is alcohol soluble and water soluble. Dissolve 0.5g of eosin in 100ml of 70% ethanol or distilled water to form a working solution;
Dyeing steps:
1. Rinse 3 times with a temperature BSS at 37 ° C for 20 s - 1 min each time;
2. Fix with neutral formalin for 30 min;
3. Rinse with distilled water once, then dilute with hematoxylin (1/20) diluted with distilled water for 10 min;
4. Rinse several times with tap water, then rinse with 1% NaHCO 3 to blue-violet;
5. Dye in the aqueous solution of eosin for 30s - 1min;
6. Rinse with distilled water and quickly pass acetone twice for 3-5 minutes each time;
7. Pass 2:1 acetone / xylene 3 times, each time 1-2 min;
8. Pass 1:2 acetone/xylene 3 times, each time 1-2 min;
9. Put in pure xylene for 5-10 minutes, observe with optical resin and observe it;
10. Place the cover slip on the slide, so that the small cover cells face up, seal with gum, and cover with a larger cover;
result:
The primary cell nuclei are stained red, and the cytoplasm is stained blue-violet;
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