H&E Staining of Mouse Tissue
From Bridges Lab Protocols
- 100% Ethanol
- 70% Ethanol
- Mayer's Hematoxylin
- 1% Eosin Y (in 95% ethanol)
- 95% Ethanol
- Removing Wax and Rehydrating
- Place slide on edge and bake for 15 min at 60 degree C to melt wax
- Deparaffinize and rehydrate sections
- Immerse for 5 min with agitation in xylene/Citrisolv
- Immerse for 5 min with agitation in 100% ethanol
- Immerse for 5 min with agitation in 70% ethanol
- Place slide(s) in appropriate buffer (For H&E, use H2O) for 1 min
- H&E staining
- Dip slide into Mayer's Hematoxylin and agitate for 30 sec
- Rinse slide in H2O for 1 minute
- depending on the intensity desired, repeat this step if necessary
- Dip slide in 1% eosin Y fo 10-30 sec with agitation
- For adipose tissue, 20 seconds allows for proper staining.
- Place slides in 95% Ethanol two times for 30 seconds each.
- Transfer slides into 100% Ethanol for 30 seconds, repeat with fresh 100% Ethanol.
- Wash twice with fresh xylene/Citrisolv for 30 seconds.
- Let slide dry.
- Place 1-2 drops of mounting medium on slide and place coverslip over
- Seal coverslip with sealant(nailpolish)
- Be sure to shake Eosin prior to placing slide in for staining because it will settle.
- The staining will get more intense after time, be sure not to oversaturate section
- Especially during the summer months, be sure to not let the slides set out overnight/extended period of time.
- Protocol take from summary of CSH Protocols; 2008; doi:10.1101/pdb.prot4987 and CSH Protocols; 2008; doi:10.1101/pdb.prot4986