Difference between revisions of "H&E Staining of Mouse Tissue"

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(Created page with "==Materials== *xylene *100% Ethanol *70% Ethanol *H2O *Mayer's Hematoxylin *1% Eosin Y (in water) *95% Ethanol ==Protocol== *Removing Wax and Rehydrating # Place slide on ed...")
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Revision as of 14:48, 27 March 2015

Materials

  • xylene
  • 100% Ethanol
  • 70% Ethanol
  • H2O
  • Mayer's Hematoxylin
  • 1% Eosin Y (in water)
  • 95% Ethanol

Protocol

  • Removing Wax and Rehydrating
  1. Place slide on edge and bake for 15 min at 60 degree C to melt wax
  2. Deparaffinize and rehydrate sections
    1. Immerse for 5 min with agitation in xylene
    2. Immerse for 5 min with agitation in 100% ethanol
    3. Immerse for 5 min with agitation in 70% ethanol
    4. Place slide(s) in appropriate buffer (For H&E, use H2O) for 1 min
  • H&E staining
  1. Dip slide into Mayer's Hematoxylin and agitate for 30 sec
  2. Rinse slide in H2O for 1 minute
    1. depending on the intensity desired, repeat this step if necessary
  3. Dip slide in 1% eosin Y fo 10-30 sec with agitation
  4. Wash twice with fresh xylene for 30 seconds.
  5. Let slide dry.
  6. Place 1-2 drops of mounting medium on slide and place coverslip over
  7. Seal coverslip with sealant(nailpolish)


Notes

  • 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


  • Protocol take from summary of CSH Protocols; 2008; doi:10.1101/pdb.prot4987 and CSH Protocols; 2008; doi:10.1101/pdb.prot4986