H&E Staining

SOP

• SOP - Flammables and Combustibles
• SOP- Irritants
• SOP- Corrosives

Rehydrate tissue sections

1. After cutting sections of tissue onto glass slides, de-pariffinize the tissue sections in xylene for 3 min. Repeat for a total of 3x 3 minute washes using fresh xylene each time
2. Rehydrate the tissue sections by immersing in 100% ethanol for 3 minutes. Repeat a second time using fresh 100% ethanol
3. Immerse sections in 95% ethanol for 3 minutes. Repeat a second time using fresh 95% ethanol
4. Immerse sections in 70% ethanol for 3 minutes
5. Rinse sections 2 to 3 times in distilled water

Stain tissue sections with haematoxylin

1. Immerse sections in pre-prepared Mayer's haematoxylin for 5 minutes
2. Rinse under running tap water (slides still in box) until water is clear. This will take about 5 minutes

Stain tissue sections with eosin, dehydrate and coverslip

1. Immerse sections in 1% eosin from 15 seconds to 1 minute
2. Rinse 3-4 times under running tap water
3. Immerse in 70% ethanol for 2 minutes
4. Immerse in in 95% ethanol for 2 minutes. Repeat a second time using fresh 95% ethanol
5. Immerse in in 100% ethanol for 2 minutes. Repeat twice more using fresh 100% ethanol
6. Immerse in xylene for 2 minutes. Repeat twice more using fresh xylene. Check staining under the microscope
7. Coverslip using Permount or other xylene-based mounting medium