EMS26920


Components:

Alcian Blue 8Gx Solution 1%, pH 2.6-3.0
Sodium Carbonate 0.3% Aqueous Solution
Periodic Acid 1% Aqueous Solution
Schiff’s Reagent
Phophomolybdic Acid-Orange G Solution


Fixation: 

After bisecting the gland and then bisecting each half, fix for 6-10 hours in Formalin

Calcium Acetate Solution followed by 4-6 hours in 70% Alcohol before

beginning dehydration and embedding.

 

Sections: 

Paraffin at 3-5µm

 

Procedure staining:

  1. Deparaffinise and hydrate to distilled water.
  2. Stain in Alcian Blue Solution for 30 minutes. Rinse briefly in water to remove excess Alcian Blue.
  3. Place for 30 minutes in Sodium Carbonate 0.3%. Wash in running water for 5 minutes.
  4. Oxidise in Periodic Acid, 1% for 10 minutes. Wash for 5 minutes in running water.
  5. Stain for 10 minutes in Schiff’s Reagent. Wash for 5 minutes in running water.
  6. Stain for 5 seconds in Phosphomolybdic Acid-Orange G Solution. Rinse briefly in water to remove excess Orange G.
  7. Dehydrate and then clear in xylene. Examine the sections while in xylene to see if the Orange G staining is too weak or too intense, and then, if necessary, rehydrate and either restain in Orange G Solution or rinse in further water.
  8. Mount with synthetic mounting medium.

 

Results:

ACTH/MSH cells - purple, red and orange-red

TSH cells - light to dark purple-blue

Gonadotropin cells - intense red, blue-purple, blue, turquoise

GH cells - orange

 

Refernces:

McManus, J.F.A. and Mowry, R. W., Staining Methods, Histologic and Histochemical, P. B. Hoeber,

NY, 1960

Clark, G., (ed.), Staining Procedures, 3rd edition, Williams & Wilkins, Baltimore, p. 166, c1973