EMS Technical Data Sheets

Sweat-Puchter Method For Amyloid

EMS Catalog #26303

Fixation

10% Buffered Neutral Formalin

Sections

Paraffin at 6 microns. 12 micron sections may be used to demonstrate minute amyloid deposits. Staining is brighter on 12 micron sections.

Staining

  1. Deparaffinize 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 Hematoxylin 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.

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.

Product Information

Sirius Red F3BA Solution
Mayer Hematoxylin
Borate Buffer
Alkaline Alcohol
Sweat-Puchter Method