Methenamin Silver Method for Argentaffin Cells
EMS Catalog #: 26481-Series
Formalin, Bouin’s (#15990-01) or other Formalin containing fixatives. Dichromates and mercury salts impair contrast between granules and background.
Cut paraffin at 3 - 5µm sections.
- Deparaffinize and hydrate slides to distilled water.
- Treat for 10 minutes in Weigert’s Iodine (#26481-01) followed by bleaching for 10 minutes in Sodium Thiosulfate, 5% (#26481-02). Wash for 10 minutes in running water. Rinse in two changes in distilled water.
- Place section in chemically clean coplin jars containing the buffered methenamine-silver nitrate solution at room temperature and place in 60°C paraffin oven for 3 – 3 ½ hours. (preheating the solution to 60°C reduces impregnation time by ½ to 1 hour). Rinse in distilled water.
- Tone for 10 minutes in Gold Chloride, 0.1% (#26481-06). Rinse in distilled water.
- Fix in Sodium Thiosulfate, 5% (#26481-02) for 2 minutes. Wash in running water for 5 minutes
- Counterstain in Acetic Safranin O, 0.1% (#26481 -07) for 5 minutes.
- Dehydrate with Acetone, clear in Xylene and mount.
|Coarse connective tissue of the submucosa||Variable amount of blackening by 3 – 31/2 hour of impregnation|
|Granules of eosinophil leukocytes, nuclei, smooth muscle & surface epithelium||Show additional blackening after incubation beyond 3 – 3 ½ hour.|
|Granules of mast cells||Remain red after nuclei & reticulum are blackened|
Clark, G, ed.: Staining Procedures, 3rd ed.: Baltimore: Williams & Wilkins Co., 1973, p. 161
Gomori, G.: Arch Path., 45:48, 1948
Burtner, H.J. and Lillie, R.D., Stain Tech., 24:225-7, 1949.