Table 4: Xylene and alcohol free H and E staining procedure using 95% lemon water as deparaffinizing agent.

Deparaffinization Diluted lemon water 95%-I At 94°C5 min
Diluted lemon water 95%-IIAt 94°C 5 min
Distilled water-IAt 94°C 5 min
Distilled water-IIAt 94°C 5 min
Wash slides in distilled waterAt 45°C30 sec
Wash slides in distilled waterAt room temperature30 sec

Neutralizing the effect of acidity in lemon waterLithium carbonate15 min
Tap water wash5 min

Nuclear stainingHarris hematoxylinAt room temperature2 min
Tap water wash2 min

DifferentiationDifferentiation in 1% acid alcoholAt room temperature1 dip

Bluing Tap water wash10 min

Cytoplasmic staining1% eosinAt room temperature1 dip
Tap water wash1 min
Wash slides in distilled water

DehydrationOver drying the sections At 60°C5 min
Approximate time required54 min