Sweat-Puchter Method For Amyloids

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:

1. Deparaffinise and hydrate to distilled water.
2. Wash in running water 5 minutes. Mordant in Buffered Neutral Formalin overnight. Wash in water for 15 minutes.
3. Alkaline Alcohol Solution for 1 hour. Rinse in distilled water for 10 seconds.
4. 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.
5. Stain in Haematoxylin Solution for five minutes. Wash in running water for at least 20 minutes to prevent fading of stain.
6. Hydrate rapidly in Absolute Alcohol. Clear in three changes of Xylene. Mount.

Stain Results:

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.