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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..."
==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 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
## 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
#Wash twice with fresh xylene for 30 seconds.
#Let slide dry.
#Place 1-2 drops of mounting medium on slide and place coverslip over
#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
*xylene
*100% Ethanol
*70% Ethanol
*H2O
*Mayer's Hematoxylin
*1% Eosin Y (in water)
*95% Ethanol
==Protocol==
*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
## 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
#Wash twice with fresh xylene for 30 seconds.
#Let slide dry.
#Place 1-2 drops of mounting medium on slide and place coverslip over
#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
