H&E Staining of Mouse Tissue: Difference between revisions
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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==Materials== | ==Materials== | ||
*xylene | *xylene/Citrisolv | ||
*100% Ethanol | *100% Ethanol | ||
*70% Ethanol | *70% Ethanol | ||
*H2O | *H2O | ||
*Mayer's Hematoxylin | *Mayer's Hematoxylin | ||
*1% Eosin Y (in | *1% Eosin Y (in 95% ethanol) | ||
*95% Ethanol | *95% 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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##depending on the intensity desired, repeat this step if necessary | ##depending on the intensity desired, repeat this step if necessary | ||
#Dip slide in 1% eosin Y fo 10-30 sec with agitation | #Dip slide in 1% eosin Y fo 10-30 sec with agitation | ||
#Wash twice with fresh xylene for 30 seconds. | ##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. | #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 | ||
#Seal coverslip with sealant(nailpolish) | #Seal coverslip with sealant(nailpolish) | ||
==Notes== | ==Notes== | ||
*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 | ||