Special Stains - Jones Methenamine Silver (JMS) Technique

The Jones Methenamine silver stain is excellent for the demonstration of glomerular and tubular basement membranes of kidney biopsies. Periodic acid formed aldehydes from certain carbohydrate containing material will selectively reduce the methenamine silver solution to metallic silver.

The ability to bind silver ions from the solution and independently reduce silver to visible metallic form is referred to as argentaffin reaction. Gold chloride is used as it tones the tissue sections and converts metallic silver to metallic gold. Unreduced silver and excess gold chloride are then removed by sodium thiosulphate. The section is then counterstained with light green.

Method

1.      Filter silver working solution (25ml distilled water, 25ml silver nitrate 10%/3% Hexamine) into a newly cleaned Coplin jar. Heat to 60°C.

2.      Take sections to water

3.      Oxidise in 1% aqueous periodic acid for 30 minutes

4.      Rinse in several changes of distilled water

5.      Place in working silver solution at 60 degrees C for 10 minutes, then check basement membranes microscopy and put back until basement membranes appear black.

6.      Wash well in several changes of distilled water

7.      Tone in 0.2% gold chloride for 5 minutes

8.      Wash in water

9.      Fix in 5% sodium thiosulphate for 5 minutes

10.Wash in water. Counterstain in light green working solution for 20 seconds

11.Wash in 1% acetic acid

12.Wash, dehydrate, clear and mount

 Results

Basement Membranes	Black
Elastic Fibres, some Collegen and Reticulin Fibres	Black
Background and other tissue	Pale green

The results can be seen below on a renal biopsy, the basement membranes are shown up clearly in black while the background is stained green to provide better contrast.

Photo – Jones Methenamine Silver x10