Puchtler-Sweat Method for Basement Membranes

EMS26356

Components:

Resorcin-Fuchsin Solution

Nuclear Fast Red Solution

Periodic Acid 0.5% Aqueous

Sodium Bisulphite Solution

Fixation:

Carnoy’s Solution

Sections:

Paraffin at 6µm

Staining Procedures:

1. Deparaffinise and hydrate to distilled water.
2. Periodic Acid, 0.5% for 5 minutes. Rinse in two changes of distilled water.
3. Sodium Bisulphite Solution for 15 hours. Rinse in five or six changes of distilled water.
4. Resorcin-Fuchsin Solution for 4 hours> Rinse in three changes of distilled water.
5. Counterstain in Nuclear Fast Red for 4 hours. Rinse in 3 changes of distilled water.
6. Dehydrate in 95% Alcohol, Absolute Alcohol, and clear in Xylene two changes each.
7. Mount with Permount or DPX.

Stain Results:

References:

Puchtler, H. and Sweat, F.: Stain Techn., 39:163, 1964.

AFIP Manual of Histological Staining Methods, 3rd ed., ED. L. Luna: New York: McGraw Hill Publications, c. 1968, p. 73