Alcian Blue
Author: Kelsey Goldman, HTL (ASCP)
Last Update: 11 July 2023
Copyright: 2023, HistologyOutlines.com
Cite this Page! -> Goldman, Kelsey. “Alcian Blue.” HistologyOutlines.Com, 11 July 2023. Accessed (today)
Special Stain
Purpose:
- To demonstrate the presence of various acidic mucins in tissue. The Alcian Blue should never stain neutral mucins. (Alcian Blue)
Control Tissues: - Small Intestine, Appendix, Colon (Alcian Blue Stain Kit (PH 2.5, Mucin Stain) (Ab150662) | Abcam)
Staining Pattern: - Alcian Blue 2.5- All Acid Mucins will stain bright blue. These include sulfomucins, sialimucins, proteoglycans
- Alcian Blue 1.0- The most acidic mucins will stain bright blue.
Reagents Needed:
Biochemistry Theory:
Sample Protocol:
Images:
Additional References:
Protocol
-Deparaffinize and rehydrate
-Add 1g of Alcian Blue 8XG to 100mL of 3% Acetic Acid. Incubate sections in this for 30 minutes
-Rinse in running tap water for 5 minutes
-Counterstain in Nuclear Fast Red for 10 minutes
-Wash in tap water for 5 minutes
clear and mount. Caution that Nuclear Fast Red will come off in alcohols. Do not put into alcohol or run quickly through.
Adapted from Suvarna, S. Kim, et al., editors. Bancroft’s Theory and Practice of Histological Techniques. Eighth edition, Elsevier, 2019.
-Deparaffinize and rehydrate
-Add 1g of Alcian Blue 8XG to 100mL of 3% Acetic Acid. Incubate sections in this for 30 minutes
-Rinse in running tap water for 5 minutes
-Counterstain in Nuclear Fast Red for 10 minutes
-Wash in tap water for 5 minutes
clear and mount. Caution that Nuclear Fast Red will come off in alcohols. Do not put into alcohol or run quickly through.
Adapted from Suvarna, S. Kim, et al., editors. Bancroft’s Theory and Practice of Histological Techniques. Eighth edition, Elsevier, 2019.
Protocol
-Deparaffinize and rehydrate
-Add 1g of Alcian Blue 8XG to 100mL of 0.1M hydrochloric Acid. Incubate sections in this for 30 minutes
-Rinse in running tap water for 5 minutes
-Counterstain in Nuclear Fast Red for 10 minutes
-Wash in tap water for 5 minutes
clear and mount. Caution that Nuclear Fast Red will come off in alcohols. Do not put into alcohol or run quickly through.
Adapted from Suvarna, S. Kim, et al., editors. Bancroft’s Theory and Practice of Histological Techniques. Eighth edition, Elsevier, 2019.
-Deparaffinize and rehydrate
-Add 1g of Alcian Blue 8XG to 100mL of 0.1M hydrochloric Acid. Incubate sections in this for 30 minutes
-Rinse in running tap water for 5 minutes
-Counterstain in Nuclear Fast Red for 10 minutes
-Wash in tap water for 5 minutes
clear and mount. Caution that Nuclear Fast Red will come off in alcohols. Do not put into alcohol or run quickly through.
Adapted from Suvarna, S. Kim, et al., editors. Bancroft’s Theory and Practice of Histological Techniques. Eighth edition, Elsevier, 2019.