Microwave Pas-Alcian Blue Method For Mucosubstance

Components:

Alcian Blue Solution

Periodic Acid, 1% Aqueous

Schiff's Reagent

Gill's Haematoxylin

Ammonia Water, 1%

Fixation:

10% buffered Neutral Formalin

Section:

Paraffin, @ 3-5µm

Staining Procedure:

1. Deparafinised and hydrate in the usual manner.
2. Place slides in a loosely covered plastic Coplin jar containing 60ml Alcian Blue Solution
3. Microwave on medium level 60 seconds. Tightly cover, agitate, and let stand 5 minutes.
4. Wash in distilled water.
5. Place slides in a loosely covered plastic Coplin jar containing 60ml Periodic Acid, 1% Aqueous.
6. Microwave on medium level 60 seconds, tighten cover, agitate, and let stand one minute.
7. Wash in distilled water.
8. Place slides in a loosely covered plastic Coplin jar containing 60ml Schiff's Reagent
9. Microwave on medium level 60 seconds, tighten cover, agitate, let stand 30 seconds.
10. Wash in warm running water for three minutes.
11. Place slides in a loosely covered Coplin jar containing 60ml Gill's Haematoxylin #2
12. Microwave on medium level for 30 seconds.
13. Immediately rinse slides in distilled water.
14. Dip once in Ammonia Water, 1% to blue haematoxylin
15. Wash slides in distilled water, dehydrate in 70%, 95% and two changes of absolute alcohol.
16. Clear in two changes Xylene and mount with Permount

Result:

Acid Mucins turquoise

Neutral Mucins magenta

Combined acid and neutral mucins purple

Nuclei blue

References:

Mathews Karen and James K. Kelly. Dept. of Histology, Foothills Hospital. Calgary , Alberta. Canada . T2N 2T9. Journal of Histotechnology Vol. 12. No 4 Dec. 1989.