Alcian Blue - PAS - Hematoxylin

STUDY
PLAY
Fixative
10% NBF or Zenker
Thickness
4-5 microns - kidney 1-3 microns
Control tissue
Kidney, cervix, mucin
Purpose
differentiate between acidic and neutral mucosubstances. Detects intestinal metaplasia
Princple
Alican stains acidic mucosubstances. PAS stains neutral mucosubstances
Reagents
1. 3% acetic acid
2. Alcian blue
3. Periodic acid
4. Schiffs reagent
5. Sodium metabisulfate
6. water
7. hematoxylin
3% acetic acid
maintain acidic state of stain
Alcian blue
stain acid mucosubstances
Periodic acid
oxidizes tissue elements to aldehydes
Schiffs reagent
stains carbohydrates
Sodium metabisulfate
removes unbound leucofuchsin
water
develops color of schiffs
hematoxylin
stains nuceli
Stains
1. acid mucosubstances
2. neutral mucosubstances
acid mucosubstances
blue
neutral mucosubstances
magenta
Error
rinse slides after staining with nuclear fast red or they become cloudy during dehyrating. Hydrate tissue properly of it will stain weakly
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