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Electron Microscopy Sciences

Technical Data Sheets

Dane's Method for Prekeratin, Keratin, & Mucin

EMS Catalog #: 26401-Series    

Fixation:

10% Buffered Neutral Formalin (#15740)

Sections:

Cut paraffin at 6µm sections.

Staining Procedures:

  1. Deparaffinize and hydrate slides to distilled water.
  2. Mayer Hematoxylin (#26401-04) for 10 minutes. Blue in running tap water for 10 minutes. Rinse in distilled water.
  3. Phloxine B 1% Solution (#26401-01) for 3 minutes. Wash in running tap water to remove excess stain. Rinse in distilled water.
  4. Alcian Blue, 0.5% (#26401-02) for 5 minutes. Wash in tap water for 2 minutes. Rinse in distilled water.
  5. Orange G Solution (#26401-03) for 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:         

Acid Mucopolysaccharides Blue
Prekeratin and Keratin Orange to Red
Nuclei Brown

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.

Online Ordering

Dane's Method for Prekeratin, Keratin, & Mucin is available online from the EMS Catalog. For ordering or product information, click here.