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H&E Staining of Mouse Tissue

207 bytes added, 14:51, 9 June 2015

→‎Protocol

==Materials==

*xylene/Citrisolv

*100% Ethanol

*70% Ethanol

*H2O

*Mayer's Hematoxylin

*1% Eosin Y (in ~~water~~95% ethanol)

*95% Ethanol

# 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

##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.

*Dehydration

#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)

==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

*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

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