Dane's Method for Prekeratin, Keratin, & Mucin

EMS26401

Components:

Phloxine Solution 1%

Alcian Blue 0.5%

Orange G Solution

Mayer's Haematoxylin

Fixation:

10% Buffered Neutral Formalin

Sections:

Cut paraffin at 6µm sections

Staining Procedures:

1. Deparaffinise and hydrate slides to distilled water.
2. Mayer Haematoxylin for 10 minutes. Blue in running tap water for 10 minutes. Rinse in distilled water.
3. Phloxine B 1% Solution for 3 minutes. Wash in running tap water to remove excess stain. Rinse in distilled water.
4. Alcian Blue, 0.5% for 5 minutes. Wash in tap water for 2 minutes. Rinse in distilled water.
5. Orange G Solutionfor 13 minutes.
6. Transfer to 95% alcohol, two changes, five dips each; absolute alcohol, two changes, fifteen dips each.
7. Clear in Xylene and mount.

Stain Results:         

References:

Dane, E.T., and Herman, D. L.: Stain Techn., 38:97, 1963

AFIP Manual of Histological Staining Methods, 3rd ed., Ed. L. Luna: NY, McGraw-Hill Publ., c. 1968, p.83.