H&E Staining

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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
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