H&E Staining
Revision as of 22:18, 13 October 2014 by Erinstephenson (Talk | contribs) (Created page with "== Rehydrate tissue sections == #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 ...")
Revision as of 22:18, 13 October 2014 by Erinstephenson (Talk | contribs) (Created page with "== Rehydrate tissue sections == #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 ...")
Rehydrate tissue sections
- 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
- Rehydrate the tissue sections by immersing in 100% ethanol for 3 minutes. Repeat a second time using fresh 100% ethanol
- Immerse sections in 95% ethanol for 3 minutes. Repeat a second time using fresh 95% ethanol
- Immerse sections in 70% ethanol for 3 minutes
- Rinse sections 2 to 3 times in distilled water
Stain tissue sections with haematoxylin
- Immerse sections in pre-prepared Mayer's haematoxylin for 5 minutes
- 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
- Immerse sections in 1% eosin from 15 seconds to 1 minute
- Rinse 3-4 times under running tap water
- Immerse in 70% ethanol for 2 minutes
- Immerse in in 95% ethanol for 2 minutes. Repeat a second time using fresh 95% ethanol
- Immerse in in 100% ethanol for 2 minutes. Repeat twice more using fresh 100% ethanol
- Immerse in xylene for 2 minutes. Repeat twice more using fresh xylene. Check staining under the microscope
- Coverslip using Permount or other xylene-based mounting medium