Protocol for H&E staining for histopathology - (Oct/26/2008 )

Hello All!!!

i need a protocol describe to me in detail how to get H&E stained sections from liver, kidney and testis for histopathological examination

which Fixtive is prefered? duration of fixation??
duration for dehydration and clearing?
also the steps for staining in hematoxylin and eosin??

Best regards

If the tissue is formalin fixed

dewax with xylene 4 times, 5 min each
dehydrate with 100% ethanol 3min once
90% ethanol 3min once,
70% ethanol 3 min twice
PBS 5 min three times
Hematoxylin 10min
H2O to rinse all the excess hematoxylin from the tank
Scott's tap water 2 min
H2O
ethanol 70% 3min
ethanol 90% 3min
ethanol 100% 3 min
xylene three times 5min each
add histomount and coverslip



Solution
Hematoxylin
Hematoxylin 1g
sodium iodate 0.2g
potassium aluminium sulphate 50g
distilled water 1000ml
allow to dissolve overnight
then add chloral hydrate 50g
citric acid 1g


Scott's Tap Water
Potassium Bicarbonate 2g
magnesium sulphate 20g
distilled water 1000ml

Go to the "Protocols" section of this site and look for the Histology section - lots of protocols there, funnily enough!