H&E Staining of Mouse Tissue: Difference between revisions
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==Materials== | ==Materials== | ||
*xylene | *xylene/Citrisolv | ||
*100% Ethanol | *100% Ethanol | ||
*70% Ethanol | *70% Ethanol | ||
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# Place slide on edge and bake for 15 min at 60 degree C to melt wax | # Place slide on edge and bake for 15 min at 60 degree C to melt wax | ||
# Deparaffinize and rehydrate sections | # Deparaffinize and rehydrate sections | ||
## Immerse for 5 min with agitation in xylene | ## 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 100% ethanol | ||
## Immerse for 5 min with agitation in 70% ethanol | ## Immerse for 5 min with agitation in 70% ethanol | ||
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#Place slides in 95% Ethanol two times for 30 seconds each. | #Place slides in 95% Ethanol two times for 30 seconds each. | ||
#Transfer slides into 100% Ethanol for 30 seconds, repeat with fresh 100% Ethanol. | #Transfer slides into 100% Ethanol for 30 seconds, repeat with fresh 100% Ethanol. | ||
#Wash twice with fresh xylene for 30 seconds. | #Wash twice with fresh xylene/Citrisolv for 30 seconds. | ||
#Let slide dry. | #Let slide dry. | ||
#Place 1-2 drops of mounting medium on slide and place coverslip over | #Place 1-2 drops of mounting medium on slide and place coverslip over | ||
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*Be sure to shake Eosin prior to placing slide in for staining because it will settle. | *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 | *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 | *Protocol take from summary of CSH Protocols; 2008; doi:10.1101/pdb.prot4987 and CSH Protocols; 2008; doi:10.1101/pdb.prot4986 | ||