EMS26303
Components:
Sirius Red F3Ba Solution 1%
Mayer's Haematoxylin
Borate Buffer, 0.1M, pH 9.0
Alkaline Alcohol
Fixation:
10% Buffered Neutral Formalin
Sections
Paraffin at 6µm. 12µm sections may be used to demonstrate minute amyloid deposits. Staining is brighter on 12µm sections.
Staining:
- Deparaffinise and hydrate to distilled water.
- Wash in running water 5 minutes. Mordant in Buffered Neutral Formalin overnight. Wash in water for 15 minutes.
- Alkaline Alcohol Solution for 1 hour. Rinse in distilled water for 10 seconds.
- Stain in preheated Sirius Red Solution for 90 minutes @ 60 °C. Rinse briefly in two changes of Borate Buffer. Wash in running water for 5 minutes.
- Stain in Haematoxylin Solution for five minutes. Wash in running water for at least 20 minutes to prevent fading of stain.
- Hydrate rapidly in Absolute Alcohol. Clear in three changes of Xylene. Mount.
Stain Results:
Amyloid | Pink to Red |
Nuclei | Blue |
Elasticia | Pink to Red |
Background | Unstained |
References:
Sweat, F. and Puchtler, H.: Arch. Path. 80:613, 1965.
AFIP Manual of Histologic Staining Methods, 3 rd edition, Ed L. Luna: New York : McGraw Hill Publications, c. 1968, p. 154.